Cholesterol-lowering effects of plant sterol esters differ in milk, yoghurt, bread and cereal.

OBJECTIVE: To measure the relative effects of each of four phytosterol ester-enriched low-fat foods (bread, breakfast cereal, milk and yoghurt) on serum lipids, plasma phytosterols and carotenoids. DESIGN: : Three research centres undertook a randomised, incomplete crossover, single-blind study consisting of four treatment periods of 3 weeks each, one of which was a control period. Each sterol-enriched test food provided 1.6 g/day of phytosterols as sterol esters. SETTING: General Community. SUBJECTS: In all 58, free-living men and women with mean age (s.d.) 54 (8) y, moderately elevated plasma total cholesterol 6.2 (0.7) mmol/l and body mass index 26.2 (3.0) kg/m(2). MAIN OUTCOME MEASURES: Serum lipids, plasma phytosterols and carotenoids. RESULTS: Serum total and LDL cholesterol levels were significantly lowered by consumption of phytosterol-enriched foods: milk (8.7 and 15.9%) and yoghurt (5.6 and 8.6%). Serum LDL cholesterol levels fell significantly by 6.5% with bread and 5.4% with cereal. They were both significantly less efficacious than sterol-enriched milk (P<0.001). Plasma sitosterol increased by 17-23% and campesterol by 48-52% with phytosterol-enriched milk and bread. Lipid-adjusted beta-carotene was lowered by 5-10% by sterols in bread and milk, respectively. CONCLUSIONS: This is the first study to demonstrate that cholesterol-lowering effects of plant sterol esters may differ according to the food matrix. Plant sterols in low-fat milk was almost three times more effective than in bread and cereal. Despite phytosterol-enriched cereal products resulting in lower serum cholesterol reductions compared to sterol-enriched milk, the detection of similar changes in plasma phytosterols demonstrated that such products still delivered and released phytosterols to the gut.

Increased small intestinal fermentation is partly responsible for the anti-nutritive activity of non-starch polysaccharides in chickens.

1. The mechanism of the anti-nutritive activities of soluble non-starch polysaccharides (NSPs) in broiler diets was investigated with emphasis on the inter-relationship between viscosity and fermentation along the gut. Isolated soluble NSP were added to a control diet to effect high gut viscosity, and in vivo depolymerisation of the NSP was achieved using a commercial glycanase. 2. Addition of soluble NSPs significantly (P < 0.01) increased gut viscosity, reduced the AME of the diet and depressed the growth and FCE of the birds. Enzyme supplementation of the NSP-enriched diet reversed the adverse effects, increasing (P < 0.01) weight gain, FCE and AME. Comparisons of the viscosities (mPa) in birds fed on the NSP-enriched diet and the same diet supplemented with enzyme were respectively: 11.9 v. 2.3 in the duodenum; 78.3 v. 4.4 in the jejunum and 409.3 v. 10.8 in the ileum. 3. Caecal volatile fatty acid concentration was markedly (P < 0.01) elevated by enzyme supplementation, whereas ileal fermentation was inhibited. 4. Microscopic examination revealed that, among birds fed on the NSP-enriched diet, there had been extensive small intestinal fermentation, which was eliminated by the enzyme supplementation. 5. Addition of a synthetic antibiotic (Amoxil) had no beneficial effects. 6. The current study demonstrated that increased fermentation occurs in the small intestine when a large amount of viscous NSPs is present in the diet and this is detrimental to the performance and well-being of poultry.

Effects of enzyme supplementation on the nutritive value of dehulled lupins.

1. Separate balance experiments were conducted to assess the potential of 2 commercial enzyme supplements to improve the nutritive value of dehulled lupin kernels. One supplement (enzyme A) contained primarily xylanase, pentosanase, hemicellulase activities and the other (enzyme B) primarily beta-glucanase, hemicellulase and pectinase activities. 2. The enzymes were added at 0, 0.25, 0.50, 0.75 and 1.00 g/kg in diets containing (g/kg) lupins 300, sorghum 543, casein 91, celite (as marker) 20, and vitamins and minerals 46. Control diets, with and without enzyme supplementation contained sorghum and casein at 800 and 134 g/kg, respectively, and no lupins. 3. Growth rates and food conversion ratios (FCR) of birds over 7 days were not affected by lupin inclusion or enzyme supplementation. FCR of broilers fed on the sorghum diet was improved by enzyme A but not by enzyme B. 4. Ileal starch digestibilities were slightly lower in birds fed on the lupin control diet (no enzyme) compared to the basal control diet. 5. Enzyme A increased the AME of the lupins from 10.01 MJ/kg DM to 11.65 MJ/kg DM when added at 0.5 g/kg. Higher rates of supplementation did not lead to further increases in AME values. 6. Enzyme A did not improve starch digestion in the diets but insoluble non-starch polysaccharides concentration in the digesta decreased (50.41-42.71 g/g acid insoluble ash marker) with increasing enzyme supplementation, suggesting that the improvement in AME was the result of increased fermentation of fibre in the hindgut. 7. Enzyme B did not affect the AME of lupins nor the ileal digestibility of nutrients, but caused an increase in the concentrations of soluble non-starch polysaccharides in the ileal digesta of chickens (19.21-35.77 mg/ml). This was accompanied by an increase in ileal digesta viscosity (11.4-34.2 m.Pa/s).

Nutritive activity of soluble rice brain arabinoxylans in broiler diets.

1. A soluble material (703 g/kg non-starch polysaccharide, 141 g/kg starch and 166 g/kg protein) of low viscosity (termed RB-NSP), was isolated in large quantities from defatted Australian rice bran using a mild alkaline extraction and ethanol precipitation. 2. The soluble non-starch polysaccharide fraction of RB-NSP comprised arabinose (0.40 mol%), xylose (0.32 mol%) galactose (0.17 mol%), glucose (0.08 mol%) and mannose (0.03 mol%). 3. RB-NSP was included at graded concentrations (0, 20, 40 and 60 g/kg) in a sorghum/casein basal diet and the diet fed to male broilers in a classical balance trial to determine apparent metabolisable energy (AME). The AME values recorded were 13.26, 13.85, 14.26 and 14.00 MJ/kg DM with a significant correlation (r = 0.65, P < 0.001) between dietary RB-NSP inclusion rate and dietary AME. 4. Feeding RB-NSP had no effect on growth, food conversion ratio or the digestibilities of starch and protein which were both high (0.98-0.99 and 0.88-0.89, respectively). 5. It was concluded that the RB-NSP may have been a substrate for hindgut fermentation in the broiler but that it possessed no anti-nutritive activity.

Non-starch polysaccharide-degrading enzymes increase the performance of broiler chickens fed wheat of low apparent metabolizable energy.

The effect of a commercial glycanase product (Avizyme TX) on the performance of 4-wk-old broiler chickens fed wheats with low and normal apparent metabolizable energy values was studied. Controls were fed a corn-based diet. Supplementation with the enzyme product significantly (P < 0.01) increased the apparent metabolizable energy of the low metabolizable energy wheat from 12.02 to 14.94 MJ/kg dry matter. The apparent metabolizable energy value of the normal wheat was increased from 14.52 to 14.83 MJ/kg dry matter; this was, however, not significant. Birds fed the low metabolizable energy wheat diet had significantly (P < 0.01) higher digesta viscosity and lower small intestinal starch and protein digestibilities than birds fed the normal wheat diet. Chickens fed the low metabolizable energy wheat tended to grow less than those fed the normal wheat diet. When the low metabolizable energy wheat+enzyme diet was fed, digesta viscosity was significantly (P < 0.01) lower (20.28 vs. 10.36 mPa.s), and small intestinal digestibility coefficient of starch was significantly (P < 0.01) greater (0.584 vs. 0.861) relative to values in birds fed the low metabolizable energy wheat diet alone. Although the protein digestibility coefficient also increased from 0.689 to 0.745, the difference was not significant. Weight gain and feed efficiency of birds fed the low metabolizable energy wheat+enzyme equaled those of controls. The enzyme product significantly (P < 0.01) increased the solubilization of non-starch polysaccharides within the gastrointestinal tract of birds fed both types of wheat diets.(ABSTRACT TRUNCATED AT 250 WORDS)

Feeding Australian Acacia gums and gum arabic leads to non-starch polysaccharide accumulation in the cecum of rats.

Exudative gums from two Australian Acacia species (A. pycnantha and A. baileyana) and gum arabic (from A. senegal) were fed to rats at graded levels (0, 20, 40, 80 g/kg), replacing cellulose in purified diets containing cholesterol plus cholic acid. Compared with consumption of the control diet containing cellulose only, consumption of the gums had no significant effects on concentrations of plasma or liver cholesterol. Plasma triacylglycerol concentrations were higher in rats fed gum arabic, whereas liver triacylglycerols were lower in rats fed the gums. The gums did not affect the total pool of volatile fatty acids in the ceca, as compared with results in controls, but did promote the relative contribution of propionate at the expense of acetate. In rats fed the diet containing cellulose (80 g/kg) the proportions of cecal acetate:propionate:butyrate were 76:15:9, whereas in the rats fed A. pycnantha gum, gum arabic and A. baileyana gum (80 g/kg) the ratios were 42:54: 4, 35:46:19 and 43:53:4, respectively. The low apparent fermentability of the gums was confirmed by the accumulation of non-starch polysaccharides in cecal digesta. In rats fed 80 g/kg A. pycnantha gum, 3.44 g of soluble non-starch polysaccharides was measured in the ceca, which was 58% of the dry weight of the cecal contents. We conclude that the biological activities of the Australian Acacia gums were similar to those of gum arabic and that these gums may have potential value as human food ingredients.

Soluble wheat pentosans exhibit different anti-nutritive activities in intact and cecectomized broiler chickens.

The role of the ceca in the anti-nutritive effect of wheat pentosans was studied in intact and cectomized broiler chickens. Addition of wheat pentosans (equivalent to 30 g pure arabinoxylans/kg diet) depressed the digestibilities of starch, protein and fatty acids in both types of birds. Cecectomized birds were less efficient (P < 0.01) in dry matter and energy utilization, but starch digestion was not influenced by cecectomy. Inclusion of isolated wheat pentosans decreased the fecal protein digestibility by 18% in intact birds and by 7% in cecectomized chickens, with the bird type x pentosan interaction being significant (P < 0.05). The ileal pentosan digestibility was not affected either by addition of isolated pentosans or by cecectomy; however, the fecal pentosan digestibility was significantly (P < 0.001) influenced. Thus, in intact birds the fecal pentosan digestibility coefficient was 0.216 in birds fed the control diet and 0.646 in those fed the diet with wheat pentosans; in cecectomized chickens the corresponding values were 0.193 and 0.399, indicating a significant influence of the hindgut microflora on pentosan digestion. The ileal and fecal digestibilities of fatty acids were also determined. There was no interaction between bird type and pentosan addition in the ileal digestibilities of fatty acids. Depressions in the fecal digestibilities of fatty acids 14:0 and 18:0 were significantly (P < 0.05) greater in intact birds. Our results indicate that anti-nutritive effects of wheat pentosans in poultry are partially due to an increased activity of hindgut microflora.

Anti-nutritive effect of wheat pentosans in broiler chickens: roles of viscosity and gut microflora.

1. The mechanism of the anti-nutritive activity of isolated wheat pentosans was investigated by examining the roles of digesta viscosity and gut microflora in broiler chickens. 2. Wheat pentosans were isolated by alkaline extraction and purified by sequential treatment with pancreatin, alpha-amylase and lichenase, and high-speed centrifugation. Some of the pentosans were depolymerised using a beta-xylanase, which reduced the relative viscosity of the polysaccharides 4 fold. 3. Inclusion of 35 g alkali-extractable pentosans (containing 854 g arabinoxylans/kg DM) per kg diet significantly (P less than 0.05) depressed broiler performance and the viscosity of the digesta of these birds was significantly (P less than 0.05) higher than that of controls. Addition of the same amount of depolymerised pentosans had no significant effect on bird performance and had less effect on digesta viscosity. 4. Supplementation of the diet containing wheat pentosans (30 g/kg) with procaine penicillin (150 mg/kg) did not improve bird performance. 4. It is concluded that the wheat pentosans elicit their anti-nutritive activity predominantly through increasing the viscosity of digesta.

The inhibition of nutrient digestion by wheat pentosans.

Two pentosan-rich fractions (water-extractable, WEP, and alkali-extractable, AEP) were isolated from a wheat milling by-product. When both WEP and AEP were added to a commercial-type broiler diet the apparent metabolizable energy (AME), nitrogen retention, feed utilization and growth of broilers were significantly (P less than 0.001) depressed. These depressions were closely correlated (P less than 0.001) to the level of pentosans in the diets. At the highest level of inclusion of pentosans (equivalent to 40 g arabinoxylans as AEP/kg) the ileal digestibilities of starch, protein and lipid were decreased by 14.6, 18.7 and 25.8% respectively.

Anti-nutritive activity of wheat pentosans in broiler diets.

1. To assess their possible anti-nutritive activity wheat pentosans were isolated from a milling by-product and added at graded levels to a sorghum-based broiler chicken diet. 2. A water-insoluble pentosan preparation (WIP, 720 g arabinoxylan/kg) caused a depression of apparent metabolisable energy (AME) of 1.63 MJ/kg DM at the highest level of inclusion (41.9 g/kg). Broilers maintained on this diet showed significant growth depression and a decrease in feed conversion efficiency. 3. A water-soluble pentosan preparation (WSP, 520 g arabinoxylan/kg) showed less anti-nutritive activity. 4. In a second experiment the AME of rice (pearled), maize, sorghum, wheat, triticale, barley and rye was determined. The AME values obtained were highly correlated (r = -0.98, P less than 0.001) with the summed levels of pentosans and beta-glucans found in the cereals. 5. It was concluded that the pentosans of wheat possess anti-nutritive activity when present in broiler diets and that similar polysaccharides may influence the nutritive value of other cereals.

Structure of the capsular polysaccharide of Escherichia coli O9:K32(A):H19.

The capsular polysaccharide of the bacterium Escherichia coli O9:K32(A):H19 was analyzed using chemical methods (hydrolysis, sequential Smith degradation, methylation analysis) together with 1H- and 13C-n.m.r. spectroscopy. 13C-N.m.r. spectroscopy and chemical analyses indicated that the K32 polysaccharide is composed of equimolar proportions of glucose, galactose, rhamnose, and glucuronic acid, and carries O-acetyl groups. 1H-N.m.r. analysis of native K32 polysaccharide revealed five resonances in the anomeric region (delta 5.52, 5.16, 5.12, 5.02, and 4.73) and the presence of an acetyl group (delta 2.18). O-Deacetylation of the polysaccharide resulted in the loss of the resonance at delta 2.18 and one of the resonances (delta 5.52) in the anomeric region. The "extra" anomeric resonance in the 1H-n.m.r. spectrum of the native K32 polymer was assigned to H-2 of rhamnose, which experiences a large downfield shift when the 2-position is O-acetylated. This was confirmed by a 2D-COSY n.m.r. experiment and studies of model compounds. The K32 capsular polysaccharide is of the "2 + 2" type, comprised of the following repeating unit: (sequence; see text) This structure is identical to that of Klebsiella K55 capsular polysaccharide.

Composition of alginate synthesized during the growth cycle of Pseudomonas aeruginosa.

A mucoid P. aeruginosa isolated from the sputum of a cystic fibrosis patient was grown in batch culture on a complex medium. During the growth cycle the amount of alginate produced was estimated and its composition was determined by proton magnetic resonance (1H-n.m.r) spectroscopy. Exopolysaccharide production occurred mainly during the exponential phase of growth. The alginate samples isolated varied little in composition and were characterized by being highly acetylated, high mannuronate (0.83-0.93 mole fraction) polymers. Guluronate was present only within heteropolymeric regions of the polysaccharides which all displayed a complete absence of polyguluronate. Ca2+ ion supplementation of the medium was not observed to increase the levels of guluronate in the alginates produced.

The detection and use of hemoglobin mutants in the direct analysis of human globin genes.

Many human globin-chain mutants contain amino acid replacements that result from single base changes in the corresponding globin gene. Using recombinants, the coding sequences of each of the alpha-, beta-, Ggamma-, and Agamma-globin genes have now been determined. Those sequences of DNA that are cleaved by a number of specific restriction endonucleases have been identified and accurately positioned. Mutations at these sequences abolish the restriction site, and therefore, the pattern of DNA fragments containing hybridizing globin-gene sequences is altered compared to DNA from normal persons. This allows the identification of one of a pair of cross-hybridizing human globin-gene sequences, as is shown here for the two alpha-globin, the two gamma-globin, and the delta- and beta-globin genes.

Model for antenatal diagnosis of beta-thalassaemia and other monogenic disorders by molecular analysis of linked DNA polymorphisms.

Polymorphisms of DNA restriction sites within the human fetal globin genes have been used to identify chromosomes that carry beta-thalassaemia genes in individuals heterozygous for this disease. This has allowed an antenatal diagnosis for beta-thalassaemia to be carried out by observation of the pattern of the inherited polymorphism of a linked DNA sequence not involved in the genetic pathogenesis of the disease. In the populations we have investigated there is no constant pattern of polymorphism that segregates with the beta-thalassaemia gene. The use of linked polymorphisms should, therefore, be applicable to antenatal diagnosis both of beta-thalassaemia and of any other single-gene defect for which there is a DNA probe specific for a sequence linked to the affected locus.

Structure of the human G gamma-A gamma-delta-beta-globin gene locus.

We have constructed a physical map of the human G gamma-, A gamma-, delta-, and beta-globin genes. The previously described maps of the fetal and adult beta-like globin genes have been linked to one another by identification of a DNA fragment, generated by BamHI, that contains part of each of the A gamma- and delta-globin genes. The map obtained, which spans more than 40 kilobases, shows the following intergene distances: between G gamma and A gamma, 3500 base pairs; between A gamma and delta, 13,500 base pairs; and between delta and beta, 5500 base pairs. All genes are transcribed from the same DNA strand.

The structure of the human beta-globin gene in beta-thalassaemia.

Twenty-one cases of beta 0 and beta +-thalassaemia have been analysed by restriction endonuclease mapping. In most cases no deletion in the regions surrounding the beta- and delta-globin genes could be detected. However, in a single Asian case of beta 0-thalassaemia, homozygous clinically, one of the homologous chromosomes contained a beta-globin gene with a deletion of 600 base pairs of DNA and comprising most or all of the 3' end of the structural gene including the EcoRI restriction site within the beta-globin coding sequence.

Structure of the human fetal globin gene locus.

We have derived a 'map' of restriction enzyme sites in and around the human gamma-globin genes. This has enabled us to show that there are two gamma-globin genes per haploid set, that the genes contain 'introns' within the same regions of DNA as the human beta and delta-globin genes, and that the genes are 3,500 base pairs apart. We conclude that the correct gene organisation of the human beta-like globin locus is GgammaAgammadeltabeta.