Arabinoxylan from Argentinian whole wheat flour promote the growth of Lactobacillus reuteri and Bifidobacterium breve.

Arabinoxylans are part of dietary fibre and have received attention given their emergent prebiotic character. Four arabinoxylans extracts were obtained from Argentinian soft and hard wheat. In vitro assays were performed to describe the extent to which the extracts from whole wheat flour support selective growth of Bifidobacterium breve and probiotic Lactobacillus reuteri ATCC23272 in a defined media. The prebiotic effect was evaluated by three quantitative scores: relative growth, prebiotic activity score and prebiotic index. For prebiotic index equation the growth of Bacteroides and Clostridium strains was compared to that of bifidobacteria and lactic acid bacteria. All the arabinoxylans extracts supported the growth of Lactobacillus and Bifidobacterium, reaching higher prebiotic activity score values than inulin (0·37 and 0·36 for Lactobacillus and Bifidobacterium respectively). AX2 from soft wheat and AX4 from hard showed similar prebiotic index value to commercial inulin (2·64, 2·52 and 2·22 respectively), and AX3 extract presented higher prebiotic index value (4·09) than the positive control and other prebiotic index reported for arabinoxylans. These extracts could be used as prebiotic, synbiotic compositions or novel food prototypes to treat dysbiosis associated with many diseases. SIGNIFICANCE AND IMPACT OF THE STUDY: The present work demonstrates that AX extracts from Argentinian soft and hard wheat promote efficiently the growth of probiotic strain L. reuteri ATCC23272 and B. breve 286, validated with three different parameters that consider the growth of representative strains of Bacteria genera found in the gut. The evaluation of AX extracts as a food supplement in a murine model could confirm their ability to modulate the microbiome. Novel food prototypes including AX and probiotics could relieve local symptoms and may act as psychobiotics with a beneficial effect on microbiome-brain axis.

A study on fibre addition to gluten free bread: its effects on bread quality and in vitro digestibility.

The aim of this study was to assess the impact of fibre addition on gluten-free (GF) dough properties and bread technological quality, and on protein and starch in vitro digestibility. Soluble (Inulin, In) and insoluble fibres (oat fibre, OF, and type IV resistant starch, RSIV) were used at 5 and 10% substitution levels. Dough firmness increased when insoluble fibres were added, and decreased when In was used. Incorporation of insoluble fibres resulted into bread with a low specific volume (SBV) since firmer dough were more difficult to expand during proofing and baking. Staling rate was reduced after fibre addition, with the exception being OF 10%, as its lower SBV may have favoured molecule re-association. In general, protein and starch digestibility increased when fibres were added at 5%, and then decreased after further increasing the level. Fibres may have disrupted bread crumb structure, thus increasing digestibility, although the higher addition may have led to a physical and/or chemical impediment to digestion. Inulin has well-known physiological effects, while RS presented the most important effect on in vitro starch digestibility (GI). These results showed the possibility of adding different fibres to GF bread to decrease the GI and increase protein digestibility, while obtaining an overall high quality end-product.

Effect of four types of dietary fiber on the technological quality of pasta.

The development of dietary fiber-enriched foods permits to obtain products with functional properties but can cause several problems in technological quality. The aim of this study was to study the quality of pasta obtained by replacing bread wheat flour with resistant starch II (RSII), resistant starch IV (RSIV), oat bran (OB) and inulin (IN) with the purpose of improving their nutritional quality. RSII, RSIV, OB and IN were substituted for a portion of bread wheat flour at levels 2.5%, 5.0%, 7.5% and 10.0%. Cooking properties, amylose and inulin losses, color and texture were measured. Finally, nutritional quality of enriched pasta was evaluated by protein losses during cooking and total dietary fiber. Microstructure of pasta was analyzed by scanning electron microscopy. Addition of RSII into pasta formulation improved the quality of the final product. RSIV-enriched pasta presented an improvement in textural characteristics and OB affected cooking properties positively up to 5% of substitution. Inulin was lost during cooking; besides, its addition negatively affected the technological quality of pasta. The results obtained in this study prove that it is possible to elaborate pasta with acceptable cooking quality and with improved nutritional characteristics by adding 10% of RSII and RSIV and 5% of OB.

Gluten-free breadmaking affected by the particle size and chemical composition of quinoa and buckwheat flour fractions.

This study aimed at assessing the effect of physicochemical properties and the particle size of different fractions of buckwheat and quinoa on the behaviour of gluten-free dough and bread quality. Quinoa and buckwheat grains were milled with a hammer mill and then separated in three fractions. These fractions where then re-milled with a cyclonic mill to obtain samples of similar sizes. Results showed that the chemical composition of these fractions was very different and played a major role on bread quality. Proteins, lipids and fibre negatively affected bread quality, whereas starch-rich fractions were more adequate for breadmaking. Re-milling quinoa and buckwheat fractions increased bread volume, although chemical composition still influenced bread properties. For hammer-milled fractions, both the finest fractions resulted in breads with higher technological quality, as well as a final product with more fibre, minerals and proteins.

Modulatory effect of steroid hormones on GnRH-induced LH secretion by cultured rat pituitary cells.

The purpose of the present experiments was to examine the short- and long-term effects of estradiol-17 beta (E2), progesterone (P), and 5 alpha-dihydrotestosterone (DHT), alone and in combination, on the gonadotrophin-releasing hormone (GnRH)-induced luteinizing hormone (LH) secretion, using an ovariectomized rat pituitary cells culture model. After 72 h in steroid-free medium, pituitary cells were further cultured for 24 h in medium with or without E2 (1 nM), P (100 nM), or DHT (10 nM). Cultures were then incubated for 5 h in the absence or presence of 1 nM GnRH with or without steroids. LH was measured in the medium and cell extract by radioimmunoassay. The results show that the steroid hormones exert opposite effects on the release of LH induced by GnRH, which seems to be dependent upon the length of time the pituitary cells have been exposed to the steroids. In fact, short-term (5 h) action of E2 resulted in a partial inhibition (64% of control) of LH release in response to GnRH, while long-term (24 h) exposure enhanced (158%) GnRH-induced LH release. Similar results were obtained with DHT, although the magnitude of the effect was lower than with E2. Conversely, P caused an acute stimulatory action (118%) on the LH released in response to GnRH and a slightly inhibitory effect (90%) after chronic treatment. GnRH-stimulated LH biosynthesis was also influenced by steroid treatment. Significant increases in total (cells plus medium) LH were observed in pituitary cells treated with E2 or DHT.(ABSTRACT TRUNCATED AT 250 WORDS)

GnRH-stimulated glycosylation (proximal and distal) of luteinizing hormone by cultured rat pituitary cells.

In the present work, the effects of GnRH on the translation (by [14C]leucine incorporation; [14C]Leu-LH) and the glycosylation of LH by rat pituitary cells in primary culture were established. The use of specific markers as radioactive precursors made it possible to discriminate the action of the neurohormone on proximal glycosylation (by[3H]mannose incorporation; [3H]Man-LH) as well as distal glycosylation (by [3H]galactose incorporation; [3H]Gal-LH) in the course of synthesis and release of LH. Pituitary cells from ovariectomized adult rats were incubated for different periods between 0 and 5 h in medium containing [14C]Leu plus [3H]Man or [14C]Leu plus [3H]Gal with or without 10 nM GnRH. GnRH increased synthesis and release of newly synthesized LH. The magnitude of the stimulatory effect on the kinetics of [14C]Leu (slope = 63.58; 158% of control) and [3H]Man (slope = 75.15; 161%) incorporation to LH was similar. The action of the neurohormone appears to be exerted on translation, the increased [3H]Man incorporation being a secondary phenomenon arising from the greater amount of available polypeptide chains as acceptors of the polymannose core. However, a direct effect of GnRH on proximal glycosylation cannot be excluded. GnRH also stimulated the kinetics of release of [14C]Leu-LH (slope = 6.14; 236% of control) and [3H]Man-LH (slope = 8.06; 191%). Comparatively, the effect of GnRH on [3H]Gal-LH was detected earlier than that on LH labeled with the other precursors; increases in rates of production (slope = 71.57; 278% of control) and release (slope = 32.08; 494%) were higher than those in [14C]Leu- and [3H]Man-LH kinetics, indicating that GnRH acts specifically on this distal step of LH glycosylation. GnRH enhanced the relative terminal glycosylation ([3H]Gal/[14C]Leu ratio) of total and release LH without modifying the relative proximal glycosylation ([3H]Man/[14C]Leu ration) of the hormone. We conclude that GnRH can induce not only changes in the quantity (greater number of molecules) but also in the quality (molecules more glycosylated) of the secreted LH by acting directly at translation and distal glycosylation level.

Kinetics of translation and glycosylation of pituitary glycoproteins.

The present study determines the basal kinetics of synthesis of translation (by [14C] leucine incorporation, [14C]leu-PROT] and of proximal (by [3H]mannose incorporation, [3H]man-PROT) and distal (by [3H]galactose incorporation, [3H]gal-PROT) glycosylation of total adenohypophyseal glycoproteins, by rat pituitary cells in primary culture. In order to obtain more information regarding the role of both steps of glycosylation on the secretory process, the effects of cycloheximide (CH; translation inhibitor) and tunicamycin (TM;glycosylation inhibitor) on the kinetics of synthesis and release of pituitary glycoproteins were also studied. Cells were incubated in medium containing [14C]leu plus [3H]man or [14C]leu plus [3H]gal, for various time-intervals (from 0.5 to 5 h) in the absence (control) or presence of different doses of CH (1.0; 4.0 or 16.0 micrograms/ml) or TM (0.5; 1.0 or 2.0 micrograms/ml). The kinetics of synthesis (slope = 3488) and release (slope = 622) of [14C]le-PROT were higher than those of the sugar precursors (slopes: [3H]man-PROT = 1751 and 526; [3H]gal-PROT = 1231 and 506). Leucine or mannose-labeled protein was barely detectable in the medium after 2 h incubation, whereas galactose-labeled protein had already been released into the incubation medium by 30 min. Cycloheximide induced translation blockage and, concomitantly, produced a marked inhibition of [3H]man incorporation. On the other hand, TM inhibited the kinetics of synthesis and release of [3H]man-PROT without affecting those of [14C]leu-PROT. The kinetics of synthesis and release of [3H]gal-PROT, although diminished, maintained linearity and increased in function of time, even in the presence of the antibiotics. Thus, the present results on glycoproteins from the pituitary gland are consistent with the previous conclusion for other mammalian glycoproteins that carbohydrate attachment occurs in several steps to molecules destined to be secreted. Addition of mannose (proximal glycosylation) is a co-translational event and that of galactose (distal glycosylation) is post-translational and can be designated as final stages in carbohydrate assembly, occurring close to the time of release. Furthermore, it has been demonstrated that the absence of the carbohydrate side chains of the pituitary glycoprotein does not prevent the intracellular transport of the protein and its export from the cell.

Rates of proximal and distal glycosylation of luteinizing hormone by cultured rat pituitary cells.

The purpose of the present work was to study the rate of basal luteinizing hormone (LH) glycosylation, discriminating the co-translational (proximal) and the post-translational (distal) glycosylation. The experiments were performed to determine the temporal relationship between the biosynthesis of the peptide chains (by [14C]leucine incorporation: [14C]Leu-LH) and the proximal (by [3H]mannose incorporation: [3H]Man-LH) and distal (by [3H]galactose incorporation: [3H]Gal-LH) glycosylation of LH, by rat pituitary cells in primary culture. In addition, the effects of cycloheximide (translation inhibitor) and tunicamycin (glycosylation inhibitor) on the rates of synthesis and release of [3H]Man-LH and [3H]Gal-LH were studied. The rates of synthesis and release (between 0 and 2 h and between 2 and 5 h) were calculated by regression analysis and the statistical significance of differences was determined by the slope test. The rate of synthesis of [3H]Man-LH (slope = 45.59) parallelled that of [14C]Leu-LH (slope = 41.39), which was in agreement with the assumption that the addition of the high mannose core is a co-translational event. Release of [3H]Man-LH (slope = 4.32) as well as that of [14C]Leu-LH (slope = 2.53) showed a lag period of approximately 2 h. The dynamics of [3H]Gal-LH secretion over the course of incubation, with a slower rate of synthesis (slope = 26.40) and a faster rate of release (slope = 6.34), differed from that of [3H]Man-LH. LH labeled with [3H]Gal was released from the early times of the incubation, indicating that galactose is added in the final stages of the secretory process into LH molecules, which are immediately released. LH translation blockage induced by cycloheximide was associated with a corresponding decrease of [3H]Man incorporation. On the other hand, addition of tunicamycin to the pituitary cells inhibited the rates of synthesis and release of [3H]Man-LH, without affecting those of [14C]Leu-LH. These findings show that proximal glycosylation depends on synthesis of the peptide chains, whereas the addition of the polymannose core is not a condition for translation. The rates of synthesis and release of [3H]Gal-LH were less affected by the antibiotics, and the inhibition was only significant at higher doses and long-time treatments. The present results demonstrate the independence of both steps of LH glycosylation, the rate of [3H]Gal-LH synthesis being 1.7-fold slower and that of release 1.5-fold faster than those of [3H]Man-LH, respectively. The data also suggest that glycosylation is not an essential step in the LH secretory process since the hormone, which is normally secreted in a glycosylated form, was synthesized, transported, and released without the carbohydrate side chains.

Effect of cycloheximide and tunicamycin on the gonadotrophin-releasing hormone stimulated distal glycosylation of luteinizing hormone by rat pituitary cells.

We have previously demonstrated that gonadotrophin-releasing hormone (GnRH) induces not only changes in quantity but also in quality on secreted luteinizing hormone (LH), by increasing [14C]Leu (translation) and [3H]Gal (distal glycosylation) incorporation into newly synthesized hormone. In the present report, we have further examined the GnRH-induced [3H]Gal-LH synthesis and release by treating anterior pituitary cells with polypeptide synthesis and glycosylation inhibitors (cycloheximide and tunicamycin, respectively). Pituitary cells from ovariectomized adult rats were cultured for 4 days and then incubated for different periods (0-5 h) in medium containing [14C]Leu plus [3H]Man or [14C]Leu plus [3H]Gal in the absence (basal) or presence of 10 nmol/L GnRH with or without (control) cycloheximide (1.0 and 4.0 microg/mL) or tunicamycin (0.5 and 2.0 microg/mL). At the end of each incubation period, the cells and the medium were separated and processed for DNA uptake and newly synthesized LH (labeled LH, by immunoprecipitation with a-betaLH) determinations. The velocity of synthesis and release (between 0 and 2 h, and between 2 and 5 h) was calculated by regression analysis and the statistical significance of differences was determined by the slope test. GnRH enhanced the rates of synthesis and release of [14C]Leu-, [3H]Man-, and [3H]Gal-LH to 157 and 237; 164 and 190; and 272 and 508% of basal values, respectively. Cycloheximide totally blocked synthesis and release of [14C]Leu-LH and greatly reduced those of [3H]Man-LH, resulting in the loss of cellular responsiveness to GnRH. Addition of tunicamycin to the pituitary cells inhibited the rates of synthesis and release of [3H]Man-LH which had been induced by GnRH, without altering those of [14C]Leu-LH. These findings indicate that glycosylation is not a condition for GnRH-stimulated LH translation. The GnRH-increased [3H]Gal-LH rates of synthesis and release were affected to a lesser extent by the inhibitors. Thus, GnRH stimulation of distal glycosylation can occur, albeit at a reduced rate, even though protein synthesis and glycosylation are blocked. In conclusion, the present results corroborate that GnRH stimulates the addition of galactose residues into LH molecule. This effect is not simply the consequence of stimulating LH polypeptide chain synthesis. In addition, it is shown that GnRH-increased LH translation is independent of glycosylation.

[Viruses precipitating asthmatic crises]. / Los virus como desencadenantes de crisis asmáticas.

Study made with 45 patients in the allergy service of the Instituto Nacional de Pediatría, with the purpose of analyzing the virus participation in the precipitation of asthmatic crisis in children who lived in the southeast of Mexico City. The subjects were included in the study if they presented signs of atopy or infection of the superior air channels. Pollen allergics, rhinosinusitis, smokers relatives and patients who own pets were excluded. In this population group, the participation of the virus were RSV, influenza B and parainfluenza.