(13)C enrichment of the CO2 in breast milk and in the breath is rapidly modified by changes in the (13)C content of the diet.

BACKGROUND: C4 plants (e.g. corn and sugar cane) have greater (13)C enrichment than C3 plants (e.g. wheat and sugar beet). OBJECTIVE: To assess whether (13)C enrichment of CO2 in the breath and breast milk of women on diets based on C3 and C4 foods changes from one diet to the other. MATERIALS AND METHODS: Six breast-feeding women were studied at 5-6 months postpartum. They ate a controlled C4 diet on days 1 and 2 followed by a C3 diet on days 3 and 4. Diet duplicates, breast milk on days 2 and 4 and hourly breath samples were collected over 4 days. (13)C enrichment was measured by isotope-ratio mass spectrometry. Values of δ(13)C were calculated from the international PDBV standard (δ(13)CPDBV). Differences between means were compared by paired t test or t test for repeated measurements. RESULTS: δ(13)CPDBV values were significantly higher in the C4 diet than in the C3 diet composites (p < 0.01). In breath CO2, the δ(13)CPDBV value was greater on days 1 and 2 (range -15.4 to -13.2, respectively) and declined on days 3 and 4 (range -20.0 to -21.8, respectively, p < 0.01). The lipid and milk serum fractions of breast milk had significantly higher δ(13)CPDBV on the C3 diet than on the C4 diet (p < 0.01). CONCLUSIONS: Subjects eating a C4 diet have a higher δ(13)CPDBV value in the breath and breast milk fractions, which diminish rapidly on a C3 diet. Further studies focusing on individual nutrients are warranted.

Effect of dietary levels of corn oil on maternal arachidonic acid synthesis and fatty acid composition in lactating rats.

OBJECTIVE: We examined the effect of different amounts of dietary corn oil rich in linoleic acid (LA) on the endogenous synthesis of arachidonic acid (AA), uptake of its precursor LA, and fatty acid composition of tissues involved in the supply of long-chain polyunsaturated fatty acids for milk synthesis. METHODS: Female Sprague Dawley rats received one of the following diets during pregnancy and lactation: a low-lipid diet (LLD; 2%), an adequate-lipid diet (ALD; 5%), or a high-lipid diet (HLD; 10%). Lipids were provided by corn oil. On day 12 of lactation we measured the endogenous synthesis of AA and quantified the conversion of (13)C-LA to (13)C-AA and the metabolic fate of (13)C-LA from all dietary groups. RESULTS: The LLD rats demonstrated larger amounts of endogenous synthesis of (13)C-AA and more dietary (13)C-LA transferred to the mammary gland (MG) than HLD rats during lactation. The proportion of medium-chain fatty acids was higher in the MG, milk clot, and liver of LLD than of HLD rats. Daily volume and 24-h yield of lipids and energy were lower in LLD rats than in HLD rats. Measurements of milk composition demonstrated that fat concentration significantly increased as lipid concentration increased in the diet. CONCLUSION: These results suggest that maternal adaptations used to compensate for diets deficient in long-chain polyunsaturated fatty acids include increased endogenous synthesis of AA and elevated uptake of LA in the MG and increased synthesis of medium-chain polyunsaturated fatty acids. It appears that the MG and liver participate together for AA synthesis for milk when this fatty acid is not provided in the diet.

Docosahexaenoic acid administered in the acute phase protects the nutritional status of septic neonates.

OBJECTIVE: We tested the hypothesis of whether a pharmacologic, orogastric dose of docosahexaenoic acid (DHA) administered during the acute phase of sepsis protects the nutritional status of neonates. METHODS: A randomized, placebo-controlled trial was conducted. Neonates who developed sepsis after a surgical procedure were randomly assigned to receive daily 100 mg of DHA or 100 mg of olive oil as placebo for 14 d. At selection, illness severity was evaluated and a blood sample was obtained to measure erythrocyte fatty acid composition. Energy intake and type of feeding were recorded daily. Body composition was also determined at selection and after 14 d of follow-up with the deuterium dilution technique. Body composition differences between d 14 and baseline were calculated and compared between groups. Confounders were analyzed in a multiple regression model. RESULTS: In all, 16 DHA and 11 placebo cases were followed up. Both groups showed an increase in length and head circumference, but length gain tended to be greater in neonates with DHA (P = 0.07). The DHA group presented increases in body mass (50 g, P = 0.03) and fat mass (70 g, P = 0.03), whereas infants in the placebo group did not show an increase in any body composition components. Gain in fat mass was positively related with the DHA of erythrocytes and whether or not infants received DHA. CONCLUSIONS: Orogastric DHA administered in the acute phase of infection likely protects the nutritional status of neonates with sepsis.

Reproducibility and reliability of the 13C-acetate breath test to measure gastric emptying of liquid meal in infants.

OBJECTIVE: This study assessed the noninvasive 13C-acetate breath test (13C-ABT) as a reliable and reproducible method to evaluate gastric emptying in infants METHODS: Gastric emptying was measured simultaneously by scintigraphy and 13C-ABT in 11 infants with clinical symptoms of gastroesophageal reflux to validate the method compared with the gold standard. Gastric emptying was also measured with 13C-ABT in a separate group of 14 healthy infants on 2 consecutive days to evaluate reproducibility of the method. Half-emptying times obtained with scintigraphy and 13C-ABT were correlated with Pearson's analysis. The variability of the half-emptying times obtained on 2 consecutive days was analyzed with paired t test and by the coefficient of variation as proposed by Bland and Altman. RESULTS: The mean emptying times obtained with scintigraphy and 13C-ABT were not different (89 +/- 27 min and 70 +/- 39 min, respectively, P = 0.22). Both methods correlated significantly (r = 0.75, P < 0.05). Half-emptying times measured on 2 consecutive days with 13C-ABT were 69 +/- 31 min and 68 +/- 30 min, respectively (mean difference = 1.4 +/- 12.4 min, P = 0.67). Intraindividual coefficient of variation was 6.3%. CONCLUSIONS: This study supports the 13C-ABT method as a useful tool to evaluate gastric emptying of liquids in healthy infants and in infants with gastroesophageal reflux in the field of research and in clinical evaluations.

[Genomic retinoblastoma perspectives: implications of tumor supressor gene RB1]. / Perspectivas en la genómica del retinoblastoma: implicaciones del gen supresor de tumor RB1.

In order to define the molecular and cellular bases of the development of retinoblastomas it is necessary to know its etiology, and to apply the advances in genome technology to this kind of neoplasia. Retinoblastomas are childhood tumors of the eye with an average incidence of one case in every 15,000-20,000 live births, which occur in sporadic and hereditary forms. The sporadic form appears regularly as a unilateral tumor, while in the familial form of the disease, tumors may be unilateral and bilateral. This neoplasia is characterized by leukocoria, strabism, and heterochromia. The retinoblastoma gene (RB1) is a molecular marker of retinoblastoma tumors. This gene is located in chromosome 13q14.2 and encodes a nuclear phosphoprotein (pRB) of 110 KDa, which plays a major role in cell proliferation control through cell cycle-regulated phosphorylation/dephosphorylation cycles of this protein. The RB1 gene is mainly affected by point mutations, which occur most frequently in exons 3, 8, 18 and 20. At the end of the last century, DNA technology has improved notably, allowing for its application to the study of a vast array of diseases. The aim of this work is to show the molecular aspects involved in retinoblastoma which are currently deciphering; this is possible thanks to new technology platforms that have been developed. This will allow us in a near future, to offer tests for the early diagnoses, prognoses, and the determination of individual predisposition towards this neoplasia.

[Molecular mechanisms of action and health benefits of polyunsaturated fatty acids]. / Mecanismos moleculares de acción de los acidos grasos poliinsaturados y sus beneficios en la salud.

Essential polyunsaturated fatty acids (PUFAs), linoleic acid n6 (LA) and linolenic acid (ALA) n3 obtained from the diet are precursors of the long-chain polyunsaturated fatty acids (Lc-PUFAs) arachidonic acid (AA) and docosahexaenoic acid (DHA) respectively. Consumption of PUFAs is related with a better neurological and cognitive development in newborns. It has been demonstrated that consumption of n-6 and n-3 PUFAs decreases blood triglycerides by increasing fatty acid oxidation through activation of PPARalpha or by reducing the activation of SREBP-1 inhibiting lipogenesis. Dietary PUFAs activate PPARalpha and PPARgamma increasing lipid oxidation, and decreasing insulin resistance leading in a reduction of hepatic steatosis. Beneficial effects of PUFAs have been observed in humans and in animals models of diabetes, obesity, cancer, and cardiovascular diseases. It is important to promote the consumption of PUFAs. Main food sources of PUFAs n-6 are corn, soy and safflower oil, and for PUFAs n-3 are fish, soy, canola oil and, flaxseed. Finally FAO/WHO recommends an optimal daily intake of n6/n3 of 5-10:1.

Synthesis of long-chain polyunsaturated fatty acids in lactating mammary gland: role of Delta5 and Delta6 desaturases, SREBP-1, PPARalpha, and PGC-1.

The purpose of this work was to study whether rat lactating mammary gland can synthesize PUFAs through the expression of Delta5 and Delta6 desaturases (Delta5D and Delta6D), whether these enzymes are regulated by the transcription factors sterol-regulatory element binding protein 1 (SREBP-1) and peroxisome proliferator-activated receptor alpha (PPARalpha) and the coactivator peroxisome proliferator-activated receptor gamma coactivator 1beta (PGC-1beta), and whether these desaturases are regulated by the lipid concentration in the diet. The results showed that on day 12 of lactation, approximately 35% of the linoleic acid in the diet, which is the precursor of PUFAs, is transferred to the mammary gland. There was expression of Delta5D and Delta6D in mammary gland, and it was regulated by the corn oil content in the diet. The higher the corn oil content in the diet, the lower the expression of both desaturases. Induction of Delta5D and Delta6D was associated positively with similar changes in SREBP-1 and PGC-1beta. Expression of PPARalpha was barely detected and was not affected by the corn oil content in the diet, whereas PGC-1beta expression increased as the corn oil in the diet increased. These results indicate that the lactating mammary gland has the capacity to synthesize PUFAs and can be regulated by the lipid content in the diet.

Perspectivas en la genómica del retinoblastoma: Implicaciones del gen supresor de tumor RB1 / Genomic retinoblastoma perspectives: Implications of supressor gene of tumor RB1

In order to define the molecular and cellular bases of the development of retinoblastomas it is necessary to know its etiology, and to apply the advances in genome technology to this kind of neoplasia. Retinoblastomas are childhood tumors of the eye with an average incidence of one case in every 15,000-20,000 live births, which occur in sporadic and hereditary forms. The sporadic form appears regularly as a unilateral tumor, while in the familial form of the disease, tumors may be unilateral and bilateral. This neoplasia is characterized by leukocoria, strabism, and heterochromia. The retinoblastoma gene (RBl) is a molecular marker of retinoblastoma tumors. This gene is located in chromosome 13q14.2 and encodes a nuclear phosphoprotein (pRB) of 110 KDa, which plays a major role in cell proliferation control through cell cycle-regulated phosphorylation/dephosphorylation cycles of this protein. The RBl gene is mainly affected by point mutations, which occur most frequently in exons 3, 8, 18 and 20. At the end of the last century, DNA technology has improved notably, allowing for its application to the study of a vast array of diseases. The aim of this work is to show the molecular aspects involved in retinoblastoma which are currently deciphering; this is possible thanks to new technology platforms that have been developed. This will allow us in a near future, to offer tests for the early diagnoses, prognoses, and the determination of individual predisposition towards this neoplasia.

El retinoblastoma es una neoplasia embrionaria que se manifiesta en dos formas: esporádica (no heredada) o familiar (heredada). En los casos esporádicos el tumor es unilateral y en la forma familiar puede presentarse de manera unilateral o bilateral. Esta neoplasia tiene una incidencia promedio de 1/15,000 nacidos vivos, presentando signos y síntomas que incluyen leucocoria, estrabismo, midriasis unilateral y heterocromía. El gen que predispone al desarrollo de retinoblastoma es RBl y se localiza en el cromosoma 13 en la región ql4.2. El gen RBl codifica para una fosfoproteína nuclear que participa de manera importante en la regulación del ciclo celular. De acuerdo con la hipótesis de Knudson, para que se desarrolle la neoplasia se deben presentar dos mutaciones en el gen RBl. Las mutaciones puntuales son las que más frecuentemente se presentan en el gen RBl; la mayoría de los estudios indican que los exones 3, 8, 18, 19 y 20 son las regiones de mutación preferencial. En la áltima década ha habido un gran avance en la tecnología del DNA, lo cual hace posible su aplicación en diferentes enfermedades. Estas herramientas moleculares podrían ser de gran utilidad en el diagnóstico o conocimiento de la predisposición a desarrollar un retinoblastoma. Entre estas valiosas herramientas se cuenta con la hibridación fluorescente realizada in situ, hibridación genómica comparativa, las microhileras y por áltimo la identificación de polimorfismos de un sólo nucleótido. En conclusión, actualmente se están descifrando los aspectos moleculares que están relacionados con el retinoblastoma, gracias a la aplicación de nuevas plataformas tecnológicas. Esto permitirá en un futuro próximo ofrecer pruebas para un diagnóstico temprano o para conocer el pronóstico y la predisposición de individuos a desarrollar esta patología. Con el fin de entender las bases celulares y moleculares del desarrollo del retinoblastoma, el objetivo del presente trabajo es mostrar el estado del arte del conocimiento de esta neoplasia, así como su origen y los avances en la genómica aplicada al retinoblastoma.

Mecanismos moleculares de acción de los ácidos grasos poliinsaturados y sus beneficios en la salud / Molecular mechanisms of action and health benefits of polyunsaturated fatty acids

Essential polyunsaturated fatty acids (PUFAs), linoleic acid n6 (LA) and linolenic acid (ALA) n3 obtained from the diet are precursors of the long-chain polyunsaturated fatty acids (Lc-PUFAs) arachidonic acid (AA) and docosahexaenoic acid (DHA) respectively. Consumption of PUFAs is related with a better neurological and cognitive development in newborns. It has been demonstrated that consumption of n-6 and n-3 PUFAs decreases blood triglycerides by increasing fatty acid oxidation through activation of PPARα or by reducing the activation of SREBP-1 inhibiting lipogenesis. Dietary PUFAs activate PPARα and PPARγ increasing lipid oxidation, and decreasing insulin resistance leading in a reduction of hepatic steatosis. Beneficial effects of PUFAs have been observed in humans and in animals models of diabetes, obesity, cancer, and cardiovascular diseases. It is important to promote the consumption of PUFAs. Main food sources of PUFAs n-6 are corn, soy and safflower oil, and for PUFAs n-3 are fish, soy, canola oil and, flaxseed. Finally FAO/WHO recommends an optimal daily intake of n6/n3 of 5-10:1.

Los ácidos grasos poliinsaturados indispensables (AGPIs), ácido linoleico n-6 y ácido linolénico n-3 se obtienen a través de la dieta y son precursores de los ácidos grasos poliinsaturados de cadena larga (AGPIs-CL) araquidónico (AA) y docosahexaenoico (ADH), respectivamente. El consumo de AGPIs está relacionado con un mejor desarrollo cerebral fetal y cognoscitivo del recién nacido. Los AGPIs pueden reducir la concentración de triacilgliceroles en la sangre a través de la oxidación de ácidos grasos por medio de la activación de PPARα o a través de la represión de SREBP-1 que inhibe la lipogénesis. El consumo de AGPIs puede ser benéfico en el control de ciertas enfermedades como la diabetes mellitus y la obesidad en la que los AGPIs activan a PPARα estimulando la oxidación de lípidos y disminuyendo la resistencia a la insulina y la esteatosis hepática. En el caso del cáncer los AGPIs pueden servir como agentes citotóxicos para ciertas células tumorales. Debido a su efecto hipolipémico y a su efecto antiinflamatorio, los AGPIs podrían tener efectos benéficos en la prevención de enfermedades cardiovasculares. Las principales fuentes alimenticias de AGPIs n-6 son los aceites de maíz, de cártamo y de soya, y las de AGPIs n-3 son la linaza y los aceites de pescados, canola y de soya. Finalmente, la FAO/ OMS recomienda un consumo óptimo de AGPIs diario en una proporción n-6: n-3 de 5-10: 1/día.

Characterization of a fluorometric method for lipoprotein lipase

A fluorometric assay for determining lipoprotein lipase (LpL) activity is described. Dibutyrilfluorescine (DBF) was used as substrate for the enzyme and the fluorescine liberated by enzymatic hydrolysis of the substrate was measured. Extracts of acetone powder from adipose tissue as an enzyme source showed characteristics of lipoprotein lipase activity, i.e., inhibition by NaCl and optimum activity in alkaline pH. There was close agreement in LPL activity when the same sample was measured simultaneously using either dibutyrilfluorescine or tri[9, 10 3H]oleylglycerol as substrate. The extent of inhibition of lipoprotein lipase by NaCl was similar with both methods. The fluorometric method detected changes in LPL activity in heart and adipose tissue realted to the nutritional status of the animal with the same specificity and sensitivity than did the radioactive method. The flluorometric method is as sensitive, less expensive and less time consuming than the radioactive method