Triacylglycerol composition in colostrum, transitional and mature human milk.

OBJECTIVE: Milk triglycerides from colostrum, transitional and mature human milk, were analyzed and compared in order to determine the differences in triacylglycerol composition throughout lactation. SETTING: Department of Food and Nutrition, University of Barcelona, Spain, and Neonatology Department of the University Hospital of Granada, Spain. SUBJECTS: Twenty-two healthy lactating women aged 21-35. DESIGN AND INTERVENTIONS: The triacylglycerol profiles of 47 breast milk samples including colostrum (1-3 days), transitional milk (7-10 days) and mature milk (25-60 days) were analyzed by high-performance liquid chromatography (HPLC), with light-scattering detection (LSD). RESULTS: Significant differences regarding several triglycerides were found between three milk classes when the Kruskal-Wallis nonparametric test was applied to 47 human milk samples that had been compared using the complete chromatographic triacylglycerol profile. The ANOVAS for each equivalent carbon number (ECN) group of triglycerides revealed significant differences between colostrum, transitional milk and mature milk. By the discriminant analysis of triacylglycerol percentages, in 19 colostrum samples, 14 transitional milk samples and 14 mature milk samples, three milk types were distinguished, and three triglycerides (peak no. 4, LnOO and SOO) were found to be the most predictive variables over all the triacylglycerol profile or ECN groups. CONCLUSIONS: Each state of lactation shows a specific profile of triacylglycerol composition in human milk. However the two most abundant triacylglycerides in colostrum, POO and POL, which account for more than 49% of the total, are also dominant in transitional (34%) and mature milk (42%).

Estradiol modulates insulin-like growth factor I receptors and binding proteins in neurons from the hypothalamus.

Trophic effects of 17 beta-estradiol (beta E2) on in vitro developing hypothalamic cells have been reported. Insulin-like growth factor I (IGF-I) is also a potent trophic factor for cultured hypothalamic cells. An interaction between sexual steroids and insulin-like growth factors (IGFs) in modulating growth of hypothalamic cells has been suggested. Thus, we tested whether beta E2 modulates the levels of IGF-I, its membrane receptor and its binding proteins in rat hypothalamic cultures. Using both neuron- and glial-enriched cultures obtained from fetal rat hypothalami we found that addition of beta E2 elicited a significant increase in IGF-I receptor levels in neurons, without affecting its affinity. On the other hand, the three different IGF-binding proteins (IGFBPs) found in the conditioned medium of the cultures were differentially modulated by beta E2 in the two types of cells studied. Overall, neuronal cultures produced greater amounts of IGFBPs after treatment with beta E2, with IGFBP2 reaching significantly higher levels. On the contrary, treatment with beta E2 did not significantly alter the amounts of IGFBPs produced by glial cells. Finally, the levels of immunoreactive IGF-I found either in the medium or in cellular extracts in both neuronal and glial cultures were not modified by treatment with beta E2. These results strongly support previous observations of a trophic synergistic interaction between IGFs and beta E2 on hypothalamic cells. Thus, an increase in IGF-I receptors and/or IGFBPs after exposure to beta E2 may result in an enhanced response of hypothalamic neurons to IGF-I.(ABSTRACT TRUNCATED AT 250 WORDS)

Basic fibroblast growth factor modulates insulin-like growth factor-I, its receptor, and its binding proteins in hypothalamic cell cultures.

Interactions between different growth factors may be important in the regulation of cell growth and differentiation in the nervous system. For instance, basic fibroblast growth factor (bFGF) regulates neuroblast division through a mechanism probably involving insulin-like growth factor-I (IGF-I). In this regard, we previously found that simultaneous addition of both factors produces an additive effect on survival and differentiation of hypothalamic neuronal and glial cells in culture. To further analyze these interactions, we explored the influence of bFGF on IGF-I, its membrane receptor, and its binding proteins in hypothalamic cells. We also tested the effects of IGF-I on its own receptor and binding proteins (IGFBPs) to determine the specificity of bFGF's actions. Treatment of neuronal and glial cultures with bFGF produced an increase in IGF-I receptors, without changing their affinity, together with an increase in the apparent M(r) of the receptor. On the other hand, IGF-I elicited a down-regulation of its own receptor in both neurons and glia, without modifying its affinity. Treatment with bFGF also produced a marked differential effect on the IGFBPs secreted by the cells. While IGFBP levels in neuronal cultures were greatly increased by bFGF, their production by glial cells was inhibited. On the other hand, IGF-I increased the amount of IGFBPs in both types of cells. Finally, addition of bFGF to the cultures elicited a dose-dependent increase in the release of IGF-I to the medium, but only a moderate increase in cellular IGF-I content, in both neurons and glia. We conclude that bFGF strongly modulates IGF-I, its receptors, and its binding proteins in the two major cell types of the hypothalamus. These findings reinforce the possibility that IGF-I and/or its receptors and binding proteins are involved in the trophic effects of bFGF on developing brain cells.

Volatile fatty acids, lactic acid, and pH in the stools of breast-fed and bottle-fed infants.

We measured fecal pH, volatile fatty acids (VFA), and lactic acid concentrations in 67 healthy infants ranging in age from 1 to 5 months. The babies were divided into five groups according to their diet: group 1: human milk, with subgroups of 1 month of age (n = 8), and 2-5 months (n = 12); group 2: formula milk, with subgroups of 1 month of age (n = 9) and 2-5 months (n = 10); group 3: cow's milk, one group of 2-5 months of age (n = 12); group 4: human milk supplemented with formula milk, 2-5 months of age (n = 8); and group 5: human milk supplemented with cow's milk, 2-5 months of age (n = 8). Fecal VFA concentration and pH of bottle-fed infants (groups 2 and 3) were significantly higher and lactic acid concentrations significantly lower than those of the breast-fed group. Groups 4 and 5 showed significantly higher pH and propionic acid concentration, and lower lactic acid concentration than Group 1 did. There were no significant differences in the data on pH, VFA, and lactic acid concentrations between the feces of formula and cow's milk groups. On the other hand, the addition of formula or cow's milk to the human milk was sufficient to alter the profile of VFA, the pH, and the lactic acid concentration of breast-fed infants stools. Low pH, low VFA, and high lactic acid concentrations make the colonic content of breast-fed babies an environment favorable for Bifidobacteria growth, and this is probably the main protective factor against gastrointestinal infections.

Estradiol promotes cell shape changes and glial fibrillary acidic protein redistribution in hypothalamic astrocytes in vitro: a neuronal-mediated effect.

We have previously shown that in hypothalamic mixed neuronal-glial cultures both astrocytic shape and distribution of glial fibrillary acidic protein (GFAP) are modified by estradiol. In the present study, we have investigated whether or not the presence of neurons is necessary for these hormonal effects. In mixed neuronal-glial hypothalamic cultures the proportion of process-bearing GFAP-immunoreactive cells was significantly increased after treatment for 30 min with 10(-12) M 17 beta estradiol. This effect was present for at least 1 day and was reverted by incubating the cells in estradiol-free medium. Estradiol incubation resulted in a progressive differentiation of GFAP-immunoreactive cells from a flattened epithelioid morphology to bipolar, radial, and stellate shapes. This effect was not observed in pure hypothalamic glial cultures. Furthermore, incubation of hypothalamic glial cells with medium conditioned by estradiol-treated mixed hypothalamic cultures did not affect the shape of GFAP-immunoreactive astrocytes. In contrast, addition of hypothalamic neurons, but not cerebellar neurons or fibroblasts, to established hypothalamic glial cultures affected the development of estradiol sensitivity in astrocytes. These results indicate that estradiol induction of shape changes in hypothalamic astrocytes is not only dependent on the presence of hypothalamic neurons, but that physical contact between astrocytes and neurons is necessary for the manifestation of the effect of this hormone.

Ontogeny of insulin-like growth factor I, its receptor, and its binding proteins in the rat hypothalamus.

A role for the insulin-like growth factors (IGFs) in brain growth and differentiation has recently been suggested. In previous studies on fetal hypothalamic cells we found a trophic influence of IGF-I on in vitro survival and differentiation of both neurons and glia. We have now investigated the expression of IGF-I, its receptor and its binding proteins in the rat hypothalamus to determine whether endogenous IGF-I might serve as a trophic factor during development of this brain area. Both IGF-I receptors and IGF-I binding proteins showed marked developmental stage-dependent variations. Thus, IGF-I receptors as measured by both binding and cross-linking techniques, were highest during fetal life and steadily decreased thereafter to reach low adult levels. Changes in receptor numbers rather than in its affinity constant accounted for the differences seen in binding activity during development. In addition, we found 3 different IGF-I binding proteins (IGFBPs) of apparent Mr of 24, 29 and 32 kDa respectively, whose levels also showed a specific developmental pattern. Highest levels of the 29 and 32 kDA IGFBPs were found in fetal and early postnatal life, whereas levels of the 24 kDa form were highest in young adults. Changes in the concentration of IGFBPs rather than in their affinities for IGF-I accounted for the different binding capacities found. Using a specific IGF-I radioimmunoassay we found that IGF-I-like immunoreactivity (IGF-I-li) levels had no direct correlation with developmental stage. IGF-I-li levels oscillated with no apparent trend throughout development of the hypothalamus.(ABSTRACT TRUNCATED AT 250 WORDS)