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Dissection of maize kernel composition and starch production by candidate gene association.

Cereal starch production forms the basis of subsistence for much of the world's human and domesticated animal populations. Starch concentration and composition in the maize (Zea mays ssp mays) kernel are complex traits controlled by many genes. In this study, an association approach was used to evaluate six maize candidate genes involved in kernel starch biosynthesis: amylose extender1 (ae1), brittle endosperm2 (bt2), shrunken1 (sh1), sh2, sugary1, and waxy1. Major kernel composition traits, such as protein, oil, and starch concentration, were assessed as well as important starch composition quality traits, including pasting properties and amylose levels. Overall, bt2, sh1, and sh2 showed significant associations for kernel composition traits, whereas ae1 and sh2 showed significant associations for starch pasting properties. ae1 and sh1 both associated with amylose levels. Additionally, haplotype analysis of sh2 suggested this gene is involved in starch viscosity properties and amylose content. Despite starch concentration being only moderately heritable for this particular panel of diverse maize inbreds, high resolution was achieved when evaluating these starch candidate genes, and diverse alleles for breeding and further molecular analysis were identified.

Base Sequence↗

Induction of secretory and serum antibody responses following oral administration of antigen with bioadhesive degradable starch microparticles.

Bioadhesive degradable starch microparticles were used to deliver antigen and immunoglobulin A (IgA)-enhancing cytokines to the oral mucosa. Degradable starch microparticle immunization groups consisted of rats dosed topically at the sublingual epithelium of the oral cavity, by subcutaneous injection in the vicinity of the major salivary glands or by oral intubation with degradable starch microparticles containing dinitrophenyl-bovine serum albumin +/- IL-5/IL-6 +/- penetration enhancer (alpha-lysophosphatidylcholine). Dinitrophenyl-bovine serum albumin was also adsorbed onto alum for salivary gland vicinity injection and administered to the oral cavity in soluble form. Animals were subjected to 3 immunization cycles, and sequential samples were assayed by radioimmunoassay for salivary IgA, tear IgA and serum IgG anti-dinitrophenyl antibodies after secondary and tertiary immunization. Salivary IgA responses were highest in degradable starch microparticle groups receiving penetration enhancer at 71 days post-secondary immunization and continued in one degradable starch microparticle((oral cavity) and two injected (salivary gland vicinity) groups for up to 88 days post-tertiary immunization. Long-term tear responses were also observed in degradable starch microparticle groups receiving penetration enhancer, but they dissipated before the salivary gland-alum responses following tertiary immunization. Serum IgG responses were most pronounced in salivary gland groups, but long-term low level responses were detectable in oral cavity groups receiving degradable starch microparticle formulations with penetration enhancer. Inclusion of IL-5 and IL-6 in oral cavity-delivered degradable starch microparticle formulations consistently enhanced tear IgA while only upregulating salivary IgA antibody responses at early time points post immunization. IL-5 and IL-6 did not enhance serum IgG antibodies in any group. These data indicate that bioadhesive degradable starch microparticles can be used as a vehicle to deliver antigen and cytokine signals to the oral cavity and, when delivered in combination with a penetration enhancer, can potentiate long-term salivary IgA responses.

Administration, Oral↗

Starch hydrolysis by Strepto-coccus equinus.

Dunican, Lawrence K. (Cornell University, Ithaca, N. Y.) and Harry W. Seeley. Starch hydrolysis by Streptococcus equinus. J. Bacteriol. 82:264-269. 1962.-In a study of starch hydrolysis by strains of Streptococcus equinus, 52 isolates were obtained and their amylolytic abilities determined. It was found that all the strains could hydrolyze starch to some extent when grown in the presence of an easily fermentable carbohydrate, viz., glucose. Without this carbohydrate the organisms did not hydrolyze starch. The hydrolysis of starch was inhibited when the organisms were grown in an atmosphere of 5% CO(2) and 95% N(2), even if grown in the presence of a fermentable monosaccharide. S. bovis, which was used as a reference organism, readily hydrolyzed starch in the absence of monosaccharides and in atmospheres containing CO(2). In no instance did S. equinus hydrolyze the starch to the level of reducing sugars. Negligible amounts of reducing sugars were recovered when the cell-free filtrates of S. equinus were incubated with starch. With S. bovis, the yield of reducing sugars under such conditions was almost quantitative. These facts extend further the differences between these related organisms. The ability to synthesize an internal starchlike polysaccharide was noted in most of the strains of S. equinus. Synthesis was found when the organisms were grown on maltose or on a starch medium containing a small amount of fermentable monosaccharide.

Bacteria↗

The structure and interactions of starch with food constituents.

For most starch-containing foods, the physical and functional properties can be traced to characteristic molecular species being present, their interactions with each other, and modifications caused by environmental conditions (moisture, temperature, shear) during processing and storage. In the present paper, the chemistry and physical chemistry of starch are discussed with an emphasis on how structure (molecular and supermolecular) and composition influence the functionality of this polysaccharide. New experimental findings brought forward on structure indicate that this polymeric carbohydrate is found in various metastable states, depending on the thermomechanical history of the product. Even more important to processing and quality attributes of starch products is the recognition that the dynamics of the supermolecular structure and interactions between starch and other food constituents are governed by the mobility of the amorphous phase of each particular system. In this respect, water, acting as a plasticizer, depresses the glass transition temperature (Tg) and thereby alters the kinetics of state transformations (e.g., gelatinization, retrogradation) and reactivity of starch. The effects of water on phase transition behavior of starch as well as the underlying molecular mechanisms to the phenomena of gelatinization, gelation, retrogradation, and starch-lipid interactions are reviewed herein. Finally, consideration is given to factors affecting the digestibility of starch from the viewpoint of processing-related changes in the susceptibility of starch to alpha-amylase.

Food Analysis↗

Starch and dietary fibre: their physiological and epidemiological interrelationships.

Until the 1980s, starch in the human diet was assumed to be totally degraded and absorbed in the small intestine. Several lines of evidence since then have indicated that this is not the case, including studies of factors controlling stool output, stool bulk on elemental diets, epidemiological studies of starch intakes in developed versus developing nations, and indications using breath hydrogen that fermentation takes place in the large intestine when starch is fed. Of the few direct estimations of starch escaping absorption in the small intestine that have been carried out, one has been conducted using intubation of healthy volunteers, where two different starch meals were fed and samples aspirated from the terminal ileum. This study demonstrated that 8-10% starch escaped absorption on average, with subjects varying from 2 to 20%. Hence a sizeable proportion of starch consumed daily may reach the large intestine, with important physiological consequences. Neither small intestinal transit time nor volume of flow were related to the extent of starch malabsorption in this study. However, many factors may play a role in the extent of malabsorption, and research is continuing to investigate physiological and food characteristics that may influence the digestion and absorption of starch.

Diet↗

Polyethylene-starch extrudates as erodible carriers for bioactive materials: I. Erodibility and in vitro dye release studies.

The objective of the present investigation is to develop and evaluate polyethylene-starch based carriers for sustaining the release of bioactive materials. Polyethylene-starch carriers were prepared by incorporating various amounts of corn starch 0, 40, 80, and 100% in polyethylene beads with and without dye. Acid orange dye was incorporated (1%) to study release patterns and the effect of starch (%) on release. The granulated mixtures were extruded to form uniform dispersions of starch and polyethylene throughout the extrudate. The extrudates obtained were studied for SEM surface characterization before, during, and after release studies to study the erosion process. In the case of 100% starch, 80% of the matrix was eroded in 5 weeks, while with 40% and 80% starch, erosion was less than 20%. The dye release studies showed that release could be sustained well over 12 weeks, depending on the starch (%) incorporated. The erosion process was studied using scanning electron microscopic techniques, which showed gradual erosion of starch particles, leaving a polyethylene skeleton. These materials show promising potential for use as carriers for bioactive materials.

Coloring Agents↗

Comparative effects of non-gelatinized corn and rice starches on the life-span of ICR mice.

We compared the effects of non-gelatinized rice and corn starches on the life-span of ICR mice. Six groups of male ICR mice consisting of 30 animals each were maintained on purified experimental diets containing either corn or rice starch and different amounts of soybean oil (6, 12 or 24%) throughout their life-time. Plots of the survival rates of the mice indicate that rice compared to corn starch conferred a longer life-span to ICR mice, although a significant difference due to the starch type was only observed in the mice fed on the 24% fat diet (p=0.012). A divergent effect of rice and corn starches on the survival rate was apparent when observations were combined with respect to the starch type regardless of the dietary fat level (p=0.005). In addition, two-way ANOVA data indicate that the mean survival time was longer for the mice given rice starch (593-645 days) than for those fed corn starch (538-580 days) (p=0.011). However, no significant difference in these parameters due to dietary fat levels was observed. The results of our study indicate that starch type is one of the determinants of longevity in mice.

Animals↗

Effects of pectin on glycaemia and insulinaemia after starch loading in normal, alloxan diabetic, and pancreatic duct-occluded rats.

In normal rats, and in rats with reduced intraluminal amylolytic activity, acute administration of low methoxyl pectin with soluble starch via an oro-gastric tube was found to delay starch digestion. In normal rats pectin slightly lowered plasma glucose levels after a starch load. In pancreatic duct-occluded rats, in which intestinal pancreatic digestive enzyme activities are markedly reduced, an increased insulin secretion and a reduced glucose response were seen after starch loading even without the addition of pectin, when compared to normal control rats. In these duct-occluded rats pectin was found to further reduce starch digestion, and markedly suppress plasma glucose levels after a starch load without changing plasma insulin levels. In moderately alloxan diabetic rats, in which intestinal amylase activity also was substantially reduced, pectin reduced the glucose response to starch to a similar extent (about 30%) as in duct-occluded rats without changing the impaired insulin response. The results suggest that pancreatic insufficiency accompanied by reduced intraluminal activities of pancreatic digestive enzyme activities may be associated with a decreased glucose and an increased insulin response to starch loading and that the ability of dietary fibre to reduce postprandial glycaemia may, inter alia, involve an inhibiting action by fibre on starch digestion, especially manifested in conditions of amylolytic insufficiency.

Amylases↗

Comparison of urea treatment with established methods of sorghum grain preservation and processing on site and extent of starch digestion by cattle.

To determine the effect of dry (D); reconstituted and ensiled (R); reconstituted and acid-treated (A); and urea-treated, high-moisture (U) sorghum grain on starch digestibility, four Angus x Hereford steers (means BW = 350 kg) with duodenal and ileal cannulas were used in a 4 x 4 Latin square design. Diets consisting of 69% ground sorghum grain were fed every 2 h in equal portions (8.2 kg/d). Diets averaged 46.5% starch and 12% CP, except for U, which averaged 14% CP due to urea treatment. Ytterbium attached to sorghum was used as a particulate marker. Duodenal, ileal, and fecal samples were taken 1 h postfeeding after a 14-d adaption to diets. Whole samples were analyzed. Preduodenal starch digestion (%) was 89, 83, 76, and 70, and starch digestion over the total tract was 99, 97, 95 and 91 for R, U, A, and D, respectively. Starch digestion proximal to each site (duodenum and ileum) was enhanced (P less than .05) by R and U compared with D. Within the small intestine, there was a linear relationship (P less than .003) between starch digestion and daily starch supply. However, digestibility of starch in the small intestine (mean = 45%) was not different among diets. Apparent digestibility of starch in the large intestine was not significantly different from digestibility in the small intestine. Urea-treated sorghum grain was equivalent to reconstituted, ensiled sorghum in digestion characteristics and was superior to dry sorghum.

Animal Feed↗

In vitro fermentation characteristics of native and processed cereal grains and potato starch using ileal chyme from dogs.

Two in vitro experiments were conducted to evaluate the ability of small intestinal bacteria of dogs to ferment native and extruded cereal grains and potato starch and cereal grain and potato flours. Substrates included barley, corn, potato, rice, sorghum, and wheat. In addition to testing native grains and flours, extruded substrates also were tested. Substrates were extruded at low temperatures (LT; 79 to 93 degrees C) and high temperatures (HT; 124 to 140 degrees C) using a Wenger extruder (model TX-52). Substrates varied widely in concentrations of rapidly digestible starch (RDS), slowly digestible starch (SDS), resistant starch (RS), and total starch (TS). Extrusion of most substrates at HT vs LT resulted in increased RDS and decreased RS concentrations. Organic matter disappearance (OMD) values attributed to microbial fermentation for a 5-h period were as high as 27% for native extruded substrates (LT potato starch) and 39% for potato flour. Average OMD was higher for cereal and potato flours than for native extruded substrates (29.9 vs 25.4%). Average molar percentages of short-chain fatty acids produced from all substrates fermented for 5 h were 73, 14, and 13% (acetate, propionate, and butyrate, respectively). Average lactate production for substrates ranked as follows: flours > native and extruded cereal grains and potato starch (0.33 and 0.18 mmol/g OM, respectively). In vitro microbial fermentation of starches by ileal bacteria can be substantial and is affected by differences in starch source, fraction, and processed form.

Animals↗

In vitro growth and starch digestion by Entodinium exiguum as influenced by the presence or absence of live bacteria.

In a preliminary study, the addition of antibiotics was shown to reduce bacterial concentrations in Entodinium exiguum cultures by more than 99% in 4 h, whereas the protozoal population was apparently unaffected. Using this procedure, the growth and amylolytic capability of Entodinium exiguum, in the presence or absence of live bacteria, was studied in vitro. Treatments for Trial 1 were protozoa plus antibiotics (PA), PA plus autoclaved bacteria (PAB), protozoa plus living bacteria (PLB), and only bacteria (BAC). Autoclaved or non-autoclaved cornstarch was used as an energy source. Treatment main effects were as follows: higher concentration of E. exiguum in PLB than in PA or PAB at 24 and 48 h (P < 0.01); PA and PAB were not different (P > 0.05); concentrations of E. exiguum higher in autoclaved cornstarch at 12 h (P < 0.05) but lower than in non-autoclaved cornstarch at 24 and 48 h (P < 0.01); and starch digestion in PLB was higher than in PA and PAB at all time periods, but only greater than BAC up to 24 h (P < 0.01). In Trial 2, only treatments PA, PLB, and BAC were tested. Rice starch and cornstarch were used as substrates. With rice starch, growth was higher in PLB than in PA at 24 and 48 h (P < 0.05). Starch digestion started earlier in PLB with rice starch (P < 0.05) but was complete for both substrates after 24 h. Up to 12 h (autoclaved cornstarch and rice starch) and 24 h (non-autoclaved cornstarch and cornstarch), the sum of digestion by bacteria and protozoa did not equal the extent of digestion in PLB, suggesting some kind of synergism. Total extent of digestion with protozoa was similar between the two sources; however, bacteria digested rice starch faster and to a greater extent than cornstarch. Approximate lag times with rice starch, autoclaved cornstarch, and non-autoclaved cornstarch were 6, 3, and 12 h for bacteria and < 6, 3, and 9 h for protozoa, respectively. Rate of digestion for non-autoclaved cornstarch was similar for bacteria and protozoa, whereas the rate of bacterial digestion was much faster with the other two substrates (autoclaved cornstarch and rice starch).

Animals↗

Effects of additional starch or fat in late-gestating high nonstarch polysaccharide diets on litter performance and glucose tolerance in sows.

The effects of feeding additional starch or fat from d 85 of gestation until parturition on litter performance and on glucose tolerance in sows that were fed a diet with a high level of fermentable nonstarch polysaccharides (NSP) were studied. The day after breeding, 141 multiparous sows were assigned to the experiment. At d 85 of gestation, sows were assigned to the treatments. Sows were fed 3.4 kg/d (as-fed basis) of a high-NSP diet or the same quantity of the high-NSP diet and an additional 360 g of starch (from wheat starch) daily, or the same quantity of the high-NSP diet and an additional 164 g of fat (from soybean oil) daily. During lactation, all sows were given free access to the same lactation diet. Approximately 1 wk before the expected time of parturition, an oral glucose tolerance test was performed in 38 randomly chosen sows by feeding pelleted glucose (3 g/kg BW0.75). Blood samples for glucose analyses were taken at -10, 10, 20, 30, 40, 50, 60, 70, 80, 90, 105, and 120 min after glucose was fed. The supply of additional dietary starch or fat did not increase piglet birth weight or total litter weight at birth. Sows that were fed the high-NSP diet had more (P = 0.097) live-born piglets and fewer (P = 0.084) stillborn piglets than did sows that were fed additional fat, whereas sows that were fed additional starch were intermediate for these variables. Piglet mortality after birth was not affected by dietary treatment. Body weight and backfat gains in the last month of gestation were higher for sows fed additional starch or fat than for sows fed the high-NSP diet (P < 0.001 and P = 0.017, respectively). Feed intake in lactation was greatest by sows fed the high-NSP diet, least by sows fed additional starch at the end of gestation, and intermediate by sows fed additional fat (P = 0.099). The differences in lactation feed intake did not result in differences in BW and backfat losses during lactation. Sows that were fed additional fat had the greatest glucose area under the curve (P = 0.044), indicating that these sows were less tolerant to glucose. In conclusion, feeding additional energy (starch or fat) in late-gestating sows that are fed a high-NSP diet did not increase litter weight at birth or piglet survival, but did increase maternal gain. Feeding sows additional energy from fat might induce glucose intolerance, whereas feeding sows additional energy from starch did not induce glucose intolerance.

Animals↗

Block-surface staining for differentiation of starch and cell walls in wheat endosperm.

A staining technique for differentiating starch granules and cell walls was developed for computer-assisted studies of starch granule distribution in cells of wheat (Triticum aestivum L.) caryopses. Blocks of embedded caryopses were sectioned, exposing the endosperm tissue, and stained with iodine potassium iodide (IKI) and Calcofluor White. Excessive tissue hydration during staining was avoided by using stains prepared in 80% ethanol and using short staining times. The IKI quenched background fluorescence which facilitated the use of higher concentrations of Calcofluor White. Cell wall definition was improved with the IKI-Calcofluor staining combination compared to Calcofluor alone. The high contrast between darkly stained starch granules and fluorescent cell walls permitted computer assisted analysis of data from selected hard and soft wheat varieties. The ratio of starch granule area to cell area was similar for both wheat classes. The starch granule sizes ranged from 2.1 microns 3 to 22,000 microns 3 with approximately 90% of the granules measuring less than 752 microns 3 (ca. 11 microns in diameter). Hard wheat samples had a greater number of small starch granules and a lower mean starch granule area compared to the soft wheat varieties tested. The starch size distribution curve was bimodal for both the hard and soft wheat varieties. Three-dimensional starch size distribution was measured for four cells near the central cheek region of a single caryopsis. The percentage of small granules was higher at the ends than at the mid-section of the cells.

Cell Wall↗

Alteration of milk fat by variation in the source and amount of starch in a total mixed diet fed to dairy cows.

The effect on milk yield and composition of the supplementation of the diets of dairy cows with wheat or potato peelings was studied at three different starch intakes (< 5, 6, and > 7.5 kg/d) for dry matter intakes around 20 kg/d. Starch supply was varied using different dietary concentrations of corn silage. Trials were conducted using Latin square designs, and all cows were fed a total mixed diet composed of corn silage, grass silage, or both; soybean meal; a mixed meal with formaldehyde treatment; minerals; and one of the following energy concentrates: potato peelings or wheat. Dry matter and energy intakes varied significantly only at the low starch concentration; higher intakes were observed when the potato peelings were fed. Body weight, milk yield, true protein content, lactose content, and protein yield were not affected by treatment. Milk fat content was higher (+3.3 g/kg) when potato peelings were fed at the high starch concentration but was unaffected at the low and medium starch concentration. Slow degradation of starch from the potato peelings in the rumen could enhance a higher delivery of precursors of milk fat synthesis in the udder. The effect on fat yield differed among starch concentrations. Milk fat content appeared to decrease for cows fed diets containing quickly degradable starch at a starch intake > 7 kg/d.

Animal Nutritional Physiological Phenomena↗

Glucose metabolism and milk yield of cows infused abomasally or ruminally with starch.

The effect of ruminal or abomasal starch infusion on milk yield and glucose metabolism of early lactation cows was measured. Four cows were continuously infused in the rumen or abomasum with partially hydrolyzed starch (1500 g/d) or were not infused (control) for three 14-d periods during wk 4 to 12 postpartum. Milk yield averaged over 40 kg/d throughout the experiment. Milk and milk lactose yields tended to increase when starch was infused and DMI was decreased, regardless of the site of infusion. Starch infusion increased mean insulin concentration and tended to decrease the concentration of serum nonesterified fatty acids. Ruminal starch infusion did not affect glucose irreversible loss rate but tended to increase glucagon concentration and decrease glucose oxidation. The increased milk yield that occurred when starch was infused ruminally relative to the milk yield of control cows could be a result of increased microbial protein supply or increased energy availability. Compared with ruminal starch infusion, abomasal starch infusion tended to increase the irreversible loss rate of glucose and to increase glucose oxidation. Abomasal infusion tended to increase plasma insulin concentration and to decrease the nonesterified fatty acid concentration relative to ruminal infusion. Infusion of starch abomasally resulted in increases of most uses of glucose, including milk lactose production, glucose oxidation, and the possible storage of glucose as body fat, which indicates that the early lactation dairy cow has a greater capacity for glucose metabolism than is provided by voluntary feed intake of average diets, but that not all available glucose is partitioned to the mammary gland. These data should be useful in testing current concepts and equations in nutritional and metabolic models of dairy cattle.

Abomasum↗

Potato and high-amylose maize starches are not equivalent producers of butyrate for the colonic mucosa.

Portal appearance of short-chain fatty acids (SCFA) produced from fermentation of three different resistant starch (RS) sources (raw potato starch, high-amylose maize starch and retrograded high-amylose maize starch) was investigated in pigs. The catheterization technique coupled with determination of portal blood flow was used to estimate SCFA uptake by the colonic mucosa. Our hypothesis was that these three RS were not equivalent butyrate providers for the colonic mucosa and that butyrate uptake would therefore be different after in vivo fermentation of each starch. The starches induced different patterns of appearance of SCFA in the portal blood; raw potato starch was the only RS source to show a significant appearance of butyrate in the portal blood. Thus, uptake of butyrate by the colonic mucosa apparently differed between starches. This finding suggests that butyrate uptake does not only depend on the flow of butyrate appearing in the lumen. Indeed, for unexplained reasons, utilization of butyrate by the colonic mucosa appeared to be less efficient when the butyrate was produced from fermentation of potato starch than when it was produced from fermentation of the other RS sources.

Amylose↗

Size characterization of barley starch granules by gravitational field-flow fractionation: a rapid, low-cost method to assess the brewing capability of different strains.

Cereal starch occurs as two types of micrometer-sized granules, large and small. Large starch granules are more susceptible to enzymatic hydrolysis. When cereal starch is used for fermentation processes, as in brewing of barley malt, the barley strains with the highest content of large starch granules should be preferred. Gravitational field-flow fractionation (GFFF) is a separation method able to fractionate starch samples at low cost and short analysis time. In this work, the search for the best GFFF conditions for the analytical separation of barley starch within an inter-laboratory approach is presented. For different barley strains cultivated under monitored conditions the size distributions of starch granules is here quickly monitored and characterized by GFFF. As a consequence, dimensional characterization of barley starch can allow for the selection of the most suitable strains with the lowest content of non-degradable starch.

Fermentation↗

Resistant starch supplementation influences blood lipid concentrations and glucose control in overweight subjects.

Resistant starch (RS) includes the sum of starch and degradation products of starch that resist small intestinal digestion and enter the colon. This study was planned to examine the effect of resistant starch on hypolipidemic actions, blood glucose, insulin levels and humoral immune responses in healthy overweight subjects. Healthy overweight subjects (over 120% of their ideal body weights) were fed either 24 g/d of resistant corn starch (RS) or regular corn starch (CS) for 21 d with their regular meals. Although this double-blind feeding regiment resulted in no significant changes in their weights or other physical parameters for the relatively acute period of intakes, there were significant lowering effects of serum total cholesterol (p < 0.05) and serum LDL-cholesterol (p < 0.05) in subjects supplemented RS. Compared with the control starch group, the RS supplementation also reduced the mean fasting serum glucose concentrations (p < 0.05). Resistant starch supplement resulted in the increase in serum immunoglobulin G (IgG) concentrations. Serum insulin and complement 3 (C3) were unaffected. Tested resistant starch supplementation was reported to be palatable with minimal bowel discomfort. These results suggest that RS supplementation improves the blood lipid profile and controls the blood glucose levels in healthy overweight subjects without bowel discomfort. Therefore, RS has a potential to be used as one of the promising food ingredients for reducing risk factors involved in the development of atherosclerosis and type 2 diabetes in overweight individuals. However, in order to prove RS as a novel therapeutic agent of cardiovascular diseases and diabetes, controlled trials with larger sample sizes and longer duration are warranted.

Adult↗