Evidence of native starch degradation with human small intestinal maltase-glucoamylase (recombinant)

Action of human small intestinal brush border carbohydrate digesting enzymes is thought to involve only final hydrolysis reactions of oligosaccharides to monosaccharides. In vitro starch digestibility assays use fungal amyloglucosidase to provide this function. In this study, recombinant N-terminal subunit enzyme of human small intestinal maltase-glucoamylase (rhMGAM-N) was used to explore digestion of native starches from different botanical sources. The susceptibilities to enzyme hydrolysis varied among the starches. The rate and extent of hydrolysis of amylomaize-5 and amylomaize-7 into glucose were greater than for other starches. Such was not observed with fungal amyloglucosidase or pancreatic alpha-amylase. The degradation of native starch granules showed a surface furrowed pattern in random, radial, or tree-like arrangements that differed substantially from the erosion patterns of amyloglucosidase or alpha-amylase. The evidence of raw starch granule degradation with rhMGAM-N indicates that pancreatic alpha-amylase hydrolysis is not a requirement for native starch digestion in the human small intestine.

Luminal substrate "brake" on mucosal maltase-glucoamylase activity regulates total rate of starch digestion to glucose

BACKGROUND: Starches are the major source of dietary glucose in weaned children and adults. However, small intestine alpha-glucogenesis by starch digestion is poorly understood due to substrate structural and chemical complexity, as well as the multiplicity of participating enzymes. Our objective was dissection of luminal and mucosal alpha-glucosidase activities participating in digestion of the soluble starch product maltodextrin (MDx). PATIENTS AND METHODS: Immunoprecipitated assays were performed on biopsy specimens and isolated enterocytes with MDx substrate. RESULTS: Mucosal sucrase-isomaltase (SI) and maltase-glucoamylase (MGAM) contributed 85% of total in vitro alpha-glucogenesis. Recombinant human pancreatic alpha-amylase alone contributed <15% of in vitro alpha-glucogenesis; however, alpha-amylase strongly amplified the mucosal alpha-glucogenic activities by preprocessing of starch to short glucose oligomer substrates. At low glucose oligomer concentrations, MGAM was 10 times more active than SI, but at higher concentrations it experienced substrate inhibition whereas SI was not affected. The in vitro results indicated that MGAM activity is inhibited by alpha-amylase digested starch product "brake" and contributes only 20% of mucosal alpha-glucogenic activity. SI contributes most of the alpha-glucogenic activity at higher oligomer substrate concentrations. CONCLUSIONS: MGAM primes and SI activity sustains and constrains prandial alpha-glucogenesis from starch oligomers at approximately 5% of the uninhibited rate. This coupled mucosal mechanism may contribute to highly efficient glucogenesis from low-starch diets and play a role in meeting the high requirement for glucose during children's brain maturation. The brake could play a constraining role on rates of glucose production from higher-starch diets consumed by an older population at risk for degenerative metabolic disorders.

Luminal starch substrate "brake" on maltase-glucoamylase activity is located within the glucoamylase subunit

The detailed mechanistic aspects for the final starch digestion process leading to effective alpha-glucogenesis by the 2 mucosal alpha-glucosidases, human sucrase-isomaltase complex (SI) and human maltase-glucoamylase (MGAM), are poorly understood. This is due to the structural complexity and vast variety of starches and their intermediate digestion products, the poorly understood enzyme-substrate interactions occurring during the digestive process, and the limited knowledge of the structure-function properties of SI and MGAM. Here we analyzed the basic catalytic properties of the N-terminal subunit of MGAM (ntMGAM) on the hydrolysis of glucan substrates and compared it with those of human native MGAM isolated by immunochemical methods. In relation to native MGAM, ntMGAM displayed slower activity against maltose to maltopentose (G5) series glucose oligomers, as well as maltodextrins and alpha-limit dextrins, and failed to show the strong substrate inhibitory "brake" effect caused by maltotriose, maltotetrose, and G5 on the native enzyme. In addition, the inhibitory constant for acarbose was 2 orders of magnitude higher for ntMGAM than for native MGAM, suggesting lower affinity and/or fewer binding configurations of the active site in the recombinant enzyme. The results strongly suggested that the C-terminal subunit of MGAM has a greater catalytic efficiency due to a higher affinity for glucan substrates and larger number of binding configurations to its active site. Our results show for the first time, to our knowledge, that the C-terminal subunit of MGAM is responsible for the MGAM peptide's "glucoamylase" activity and is the location of the substrate inhibitory brake. In contrast, the membrane-bound ntMGAM subunit contains the poorly inhibitable "maltase" activity of the internally duplicated enzyme.

Comparison of modal analysis results of laser vibrometry and nearfield acoustical holography measurements of an aluminum plate

Noise and vibration has long been sought to be reduced in major industries: automotive, aerospace and marine to name a few. Products must be tested and pass certain levels of federally regulated standards before entering the market. Vibration measurements are commonly acquired using accelerometers; however limitations of this method create a need for alternative solutions. Two methods for non-contact vibration measurements are compared: Laser Vibrometry, which directly measures the surface velocity of the aluminum plate, and Nearfield Acoustic Holography (NAH), which measures sound pressure in the nearfield, and using Green’s Functions, reconstructs the surface velocity at the plate. The surface velocity from each method is then used in modal analysis to determine the comparability of frequency, damping and mode shapes. Frequency and mode shapes are also compared to an FEA model. Laser Vibrometry is a proven, direct method for determining surface velocity and subsequently calculating modal analysis results. NAH is an effective method in locating noise sources, especially those that are not well separated spatially. Little work has been done in incorporating NAH into modal analysis.

High cycle fatigue of AA6082 and AA6063 aluminum extrusions

The high cycle fatigue behavior of hollow extruded AA6082 and AA6063 aluminum extrusions has been studied. Hollow extruded aluminum profiles can be processed into intricate shapes, and may be suitable replacements for fatigue critical automotive applications requiring reduced weight. There are several features inherent in hollow aluminum extrusions, such as seam welds, charge welds, microstructural variations and die lines. The effects of such extrusion variables on high cycle fatigue properties were studied by taking specimens from an actual car bumper extrusion. It appears that extrusion die lines create large anisotropy differences in fatigue properties, while welds themselves have little effect on fatigue lives. Removal of die lines greatly increased fatigue properties of AA6082 specimens taken transverse to the extrusion direction. Without die lines, anisotropy in fatigue properties between AA6082 specimens taken longitudinal and transverse to the extrusion direction, was significantly reduced, and properties associated with the orientation of the microstructure appears to be isotropic. A fibrous microstructure for AA6082 specimens showed great improvements in fatigue behavior. The effects of elevated temperatures and exposure of specimens to NaCl solutions was also studied. Exposure to the salt solution greatly reduced the fatigue lives of specimens, while elevated temperatures showed more moderate reductions in fatigue lives.

High temperature, low cycle fatigue of a hybrid particulate/fiber aluminum metal-matrix composite

The effect of shot particles on the high temperature, low cycle fatigue of a hybrid fiber/particulate metal-matrix composite (MMC) was studied. Two hybrid composites with the general composition A356/35%SiC particle/5%Fiber (one without shot) were tested. It was found that shot particles acting as stress concentrators had little effect on the fatigue performance. It appears that fibers with a high silica content were more likely to debond from the matrix. Final failure of the composite was found to occur preferentially in the matrix. SiC particles fracture progressively during fatigue testing, leading to higher stress in the matrix, and final failure by matrix overload. A continuum mechanics based model was developed to predict failure in fatigue based on the tensile properties of the matrix and particles. By accounting for matrix yielding and recovery, composite creep and particle strength distribution, failure of the composite was predicted.

Processing and mechanical properties of cast aluminum containing scandium, zirconium, and ytterbium

A series of aluminum alloys containing additions of scandium, zirconium, and ytterbium were cast to evaluate the effect of partial ytterbium substitution for scandium on tensile behavior. Due to the high price of scandium, a crucible-melt interaction study was performed to ensure no scandium was lost in graphite, alumina, magnesia, or zirconia crucibles after holding a liquid Al-Sc master alloy for 8 hours at 900 °C in an argon atmosphere. The alloys were subjected to an isochronal aging treatment and tested for conductivity and Vickers microhardness after each increment. For scandium-containing alloys, peak hardnesses of 520-790 MPa, and peak tensile stresses of 138-234 MPa were observed after aging from 150-350 °C for 3 hours in increments of 50 °C, and for alloys without scandium, peak hardnesses of 217-335 MPa and peak tensile stresses of 45-63 MPa were observed after a 3 hour, 150 °C aging treatment. The hardness and tensile strength of the ytterbium containing alloy was found to be lower than in the alloy with no ytterbium substitution.

SOLUTE-DERIVED THERMAL STABILITY OF NANOCRYSTALLINE ALUMINUM AND PROCESSING FACTOR INFLUENCE ON THE FORMATION OF AL6MN QUASICRYSTALS IN MELT-SPINNING

Thermal stability of nanograined metals can be difficult to attain due to the large driving force for grain growth that arises from the significant boundary area constituted by the nanostructure. Kinetic approaches for stabilization of the nanostructure effective at low homologous temperatures often fail at higher homologous temperatures. Thermodynamic approaches for thermal stabilization may offer higher temperature stability. In this research, modest alloying of aluminum with solute (1 at.% Sc, Yb, or Sr) was examined as a means to thermodynamically stabilize a bulk nanostructure at elevated temperatures. After using melt-spinning and ball-milling to create an extended solid-solution and nanostructure with average grain size on the order of 30-45 nm, 1 h annealing treatments at 673 K (0.72 Tm) , 773 K (0.83 Tm) , and 873 K (0.94 Tm) were applied. The alloys remain nanocrystalline (<100 nm) as measured by Warren-Averbach Fourier analysis of x-ray diffraction peaks and direct observation of TEM dark field micrographs, with the efficacy of stabilization: Sr>Yb>Sc. Disappearance of intermetallic phases in the Sr and Yb alloys in the x-ray diffraction spectra are observed to occur coincident with the stabilization after annealing, suggesting that precipitates dissolve and the boundaries are enriched with solute. Melt-spinning has also been shown to be an effective process to produce a class of ordered, but non-periodic crystals called quasicrystals. However, many of the factors related to the creation of the quasicrystals through melt-spinning are not optimized for specific chemistries and alloy systems. In a related but separate aspect of this research, meltspinning was utilized to create metastable quasicrystalline Al6Mn in an α-Al matrix through rapid solidification of Al-8Mn (by mol) and Al-10Mn (by mol) alloys. Wheel speed of the melt-spinning wheel and orifice diameter of the tube reservoir were varied to determine their effect on the resulting volume proportions of the resultant phases using integrated areas of collected x-ray diffraction spectra. The data were then used to extrapolate parameters for the Al-10Mn alloy which consistently produced Al6Mn quasicrystal with almost complete suppression of the equilibrium Al6Mn orthorhombic phase.

The Electrolytic Deposition of Chromium Upon Aluminum

The purpose or this investigation is primarily to determine the best conditions for plating chromium on aluminum. The work was carried out with the hope of obtaining coherent deposits, and of determin­ing the conditions under which such deposits may be duplicated.

The Rate of Precipitation of Copper Aluminide in the Silver Rich Silver-Copper-Aluminum Alloys

In order to determine the best annealing temperature at which to age-harden the alloys, hardness tests on speci­men annealed for different lengths of time at different temperatures were made.

The Electrical Conductivity of the Copper-Aluminum Alloys.

Among the many aluminum alloys which have been studied are the binary copper-aluminum alloys. These have proven to be among the most useful of the alumi­num alloys thus far worked upon.

The Age-Hardening of Magnesium with Aluminum and Zinc

Magnesium is one of the most active elements and forms oxides, nitrides, and carbides, but not hydrides. Due to its activity, low melting point, low strength when unalloyed, and the difficulty with which it is worked, magnesium has not been and is not at present well developed.

The Age-Hardening of Aluminum with MgZn2

The purpose of this thesis is to investigate the age-hardening of aluminum with magnesium and zinc in such proportions as to conform to the compound MgZn2. Because of a lack of time and proper equipment, the only property investigated was the hardness as indicated by the Rockwell Superficial Hardness Tester.

The Determination of Aluminum in the Presence of Manganese and Iron by the Use of a Mercury Cathode

Since 1880, when Wolcott Gibbs made the suggestion that mercury could be used as a cathode in gravimetric electroanalysis, many articles have appeared in literature either criticizing the method or citing successful results which have been obtained by it.

Age Hardening of Copper-Aluminum Alloy Castings

Age hardening occurs in alloys of the solid solution type containing a hardening constituent, be it metal or metallic compound, which is more soluble in the solvent phase at higher temperatures than at lower ones.