Standardized Ileal Amino Acid Digestibility of Corn, Corn Distillers' Dried Grains with Solubles, Wheat Middlings, and Bakery By-Products in Broilers and Laying Hens.

Standardized ileal amino acid digestibility (SIAAD) of 5 samples of corn distillers dried grain with solubles (DDGS), 5 samples of bakery by-products (BBP), 3 samples of corn, and 1 sample of wheat middlings (WM) were evaluated in broilers and laying hens. Diets containing each of the 14 feed ingredients were evaluated in 21 day-old broiler chickens. The DDGS and BBP containing diets were fed to 30-week-old laying hens, while corn and wheat middling were evaluated in 50-week-old laying hens. All the diets were semi-purified with each feed ingredient being the only source of amino acid (AA). To obtain SIAAD values, apparent ileal AA digestibility was corrected for basal ileal endogenous AA losses using values generated from broilers and laying hens fed a nitrogen-free diet. Ileal crude protein digestibility for the 5 DDGS samples was higher (P < 0.05) in broilers than in laying hens. Broilers had higher SIAAD for DDGS 2, 3, 4, and 5 while there was no difference for DDGS 1 except for 4 AA where broilers had higher (P < 0.05) SIAAD values. Standardized ileal AA digestibility values for broilers were higher (P < 0.05) for BBP 1 and 4. Ileal CP digestibility for corn 1 was higher (P < 0.05) for broilers compared to laying hens, and SIAAD values for the 16 AA (9 indispensable and 7 dispensable) evaluated in this study were higher (P < 0.05) in broilers. Broilers had higher (P < 0.05) SIAAD values for 4 (histidine, leucine, phenylalanine, and valine) and 6 (histidine, leucine, methionine, phenylalanine, threonine, and valine) indispensable and 3 (cysteine, glutamic acid, and proline) and 4 (cysteine, glutamic acid, proline, and serine) dispensable AA for corn 2 and corn 3, respectively. No difference in SIAAD between broilers and laying hens was observed for WM. Results from this study confirm that high variability in digestibility exists between different samples of DDGS. Differences in SIAAD between broilers and laying hens were observed in some samples of DDGS and BBP.

Standardized ileal amino acid digestibility of meat and bone meal and soybean meal in laying hens and broilers.

The objective of this study was to determine the standardized ileal amino acid digestibility (SIAAD) of 7 meat and bone meal (MBM) and 3 soybean meal (SBM) samples in broilers (Ross 708) and laying hens (Hy-line W36). All 10 feed ingredients were evaluated in 21-d-old broiler chickens and 30- or 50-wk-old laying hens. Standardization was accomplished by correcting for basal ileal endogenous amino acid losses using a nitrogen-free diet. Broilers were reared in cages from d 0 to 16 on a standard broiler starter diet adequate in all nutrients and energy; thereafter, they were allotted to treatments using a randomized complete design with 6 replicate cages of 8 birds each. For the laying hens, 6 replicate cages of 6 birds each (542 cm(2)/bird) were used. Each treatment diet, which was fed for 5 d, was semipurified, with MBM or SBM being the sole source of amino acids in each diet. Ileal endogenous amino acid losses were not different between broilers and the 2 groups of laying hens. Meat and bone meal from different locations varied widely in digestibility. Broilers had higher (P < 0.05) SIAAD in 4 of the 7 MBM samples. In 2 of the 3 SBM samples, broilers had higher (P < 0.05) SIAAD for most of the nonessential amino acids. Generally, hens had 6.4 and 7.7% units less Met and Lys digestibility of all MBM samples after standardization. Dry matter digestibility values of the SBM samples were higher (P < 0.05) in broilers. Likewise, broilers had 4.1 and 1.5% units more Met and Lys digestibility of all the SBM samples evaluated compared with those from laying hens. The results of these experiments suggest that differences exist in the digestive capabilities of laying hens and broilers, which indicates that species-specific nutrient digestibility values or adjustments may be needed.

The effects of copper on the efficacy of phytase, growth, and phosphorus retention in broiler chicks.

Copper is often added to broiler diets at prophylactic concentrations as an antimicrobial despite purported chelation with and reduced utilization of phytin phosphorus. Therefore, male chicks were fed 0, 62.5, 125, 250, or 375 ppm Cu from Cu sulfate in combination with 600 phytase units (FTU)/kg phytase from 9 to 22 d of age (6 cages/diet, 8 birds/cage). Nonphytate phosphorus (NPP) and Ca were formulated to 0.2 and 0.7% of the diet, respectively. Three additional control diets were formulated to contain 0.27, 0.34, and 0.40% NPP, each with 0.7% Ca. Birds fed increasing concentrations of Cu with 600 FTU phytase/kg had linear reductions in performance characteristics (P < or = 0.05). Birds fed increasing concentrations of Cu with 600 FTU phytase/kg had linear increases in toe ash percentage (P < or = 0.027), but tibia ash percentage was not affected (P > 0.05). Birds fed increasing Cu concentrations with 600 FTU phytase/kg had linear reductions in apparent P retention as a percentage of total P (P < or = 0.0005). Supplementation with increasing concentrations of Cu to a diet containing 600 FTU phytase/kg resulted in decreases in 21 d BW, BW gain, feed consumption, feed conversion, tibia and toe ash weights, and apparent P retention as a percentage of total P. In this experiment, Cu supplementation did not reduce the efficacy of phytase (i.e., improvement in apparent P retention with phytase supplementation) but did decrease apparent P retention, BW gain, feed consumption, feed conversion, and tibia ash and toe ash weights.

The selective suppression of immunogenicity by hyaluronic acid.

A hyaluronidase-sensitive component of human peritoneal fluid from a patient with Wilms' tumor when injected into rabbits has been shown to suppress the formation of humoral precipitating antibodies to certain major classes of proteins present in the fluid. Furthermore, it has been found that hyaluronic acid, when included with certain test antigens (serum albumin, fetuin) or antigen mixtures (tumor isolates or mixtures of albumin, immunoglobulin G and immunoglobulin M), produces a marked distortion or complete blockage of immunoelectrophoresis precipitin arcs, as well as altered gel chromatography elution profiles. These findings that hyaluronic acid can interfere profoundly with both the elicitation of a complete antibody response and the formation of "normal" patterns of antigen-antibody precipitates in laboratory tests supports the possibility that this polysaccharide may play an immuno-regulatory role by masking potential immunogens. Consideration of the mechanisms for these in vivo and in vitro effects suggests that there may be some common basis in an "excluded volume" property of the hyaluronate, but this does not appear sufficient to explain the complexity and selectivity of the observed phenomena.

Determination of vitamin E in microsamples of serum by liquid chromatography with electrochemical detection.

In this procedure for determination of vitamin E by "high-performance" liquid chromatography with electrochemical detection, 25-microL serum specimens are deproteinized with ethanol. Vitamin E (alpha-tocopherol), its derivatives (beta- and gamma-tocopherols), and the internal standard (delta-tocopherol) are extracted into heptane and the extract is evaporated and the residue reconstituted with methanol before injection into the chromatograph. Within- and between-run CVs for an alpha-tocopherol concentration of 13.6 mg/L were 5.1% (n = 28) and 6.0% (n = 5), respectively. The standard curve is linear to 100 mg/L; the minimum concentration detectable is 0.1 mg/L. Analytical recovery ranged from 99.8% to 104.8%. In 36 specimens collected from apparently healthy subjects who were not taking vitamin supplements, alpha-tocopherol as determined by this method ranged from 4.3 to 9.7 mg/L, from 1.8 to 3.9 mg/L for beta- and gamma-tocopherols. Results by this method (y) and an HPLC-ultraviolet method (x) correlate reasonably (r = 0.81): y = 0.88x - 0.55 mg/L (n = 45). This procedure is adaptable to automated analysis, and the small sample requirement facilitates its applicability to neonates.

Evaluation of a mini-column chromatographic procedure for the measurement of hemoglobin A1c.

An ion-exchange chromatographic procedure is described which facilitates the determination of beta-chain aminoterminal modified glycated hemoglobin. The procedure includes an erythrocyte lysis reagent which eliminates the labile aldimine component (pre-A1c) and a two-stage elution step which separates HbA, 1a + b from HbA1c. This procedure also includes calibrator material which aids in correcting for temperature fluctuations during the analysis. Within-run CV's for samples with HbA1c levels between 4.0% and 13.7% were 1.4 to 3.2%. The between-run CV for an HbA1c control was 5.5%. A comparison of the present test to an ion-exchange HPLC method yielded the equation: HPLC = 0.96 (present method) -0.2% (n = 101 and r = 0.984). Two separate reference range studies yielded comparable results (n = 220/65, mean = 4.77/4.78%, S.D. = 0.68/0.55). Studies with pooled erythrocytes and various lipemic plasmas did not reveal any assay interferences. Various abnormal hemoglobins were studied for their effect on the assay.

Glycosaminoglycan in the blood and renal tissue of a patient with nephroblastomatosis.

Bilateral nephroblastomatosis was diagnosed in a 15-month-old white female. Prior to surgery, multiple peripheral blood smears (Wrights' stain) revealed an azurophilic staining extracellular material. When serum was added to a three percent acetic acid solution, a floccular, fibrous precipitate formed at the meniscus of the tube. Serum protein electrophoresis on cellulose acetate support media resulted in a distorted pattern which corrected to a normal pattern upon treatment with hyaluronidase. These peripheral blood abnormalities disappeared following a left nephrectomy. Quantitative chemical analysis of diseased renal tissue yielded 81 micrograms of readily extracted glycosaminoglycan (GAG) per gram of tissue. The importance of abnormal glycosaminoglycan production in patients with malignant disease is discussed both in terms of clinical importance and possible roles of cell exudates.

Determination of bismuth in urine by atomic absorption with hydride generation.

A procedure for the determination of urinary bismuth by atomic absorption spectroscopy with hydride generation was developed and evaluated. Specimen or standard solutions were mixed with an acid mixture and an antifoam reagent. Sodium borohydride solution was then introduced to the reaction flask in order to produce bismuth hydride. The preliminary reference range for urinary bismuth was found to be less than 17 micrograms/L in 20 healthy control subjects. For patients on medications or medical treatments, bismuth levels varied from 5 to 1,460 micrograms/L. The minimum detection limit was found to be 2.5 micrograms/L and the procedure was linear to 250 micrograms/L. The intra-assay and interassay coefficients of variation at the level of 21 micrograms/L were 4.0 (N = 33) and 4.1% (N = 19), respectively. Average bismuth recovery was 97.7% for concentrations ranging from 25 to 100 micrograms/L. This procedure is simple, fast, and sensitive enough to detect levels well into the reference range. Preliminary studies also indicate this method can be used for serum bismuth determinations.

Centrifugal analysis for serum iron and iron-binding capacity using the IL Multistat III Analyzer.

A colorimetric method for the determination of serum iron and total iron-binding capacity (TIBC) has been adapted to an IL Multistat III Centrifugal Analyzer. The correlation to an ESA Ferrochem, Model 3050, which measures iron by coulometry, is acceptable (IL Multistat = 1.08 ESA--5.05). The intra-run precision gave an average C.V. of less than 5%. The day-to-day precision was between 2% and 7% and the average analytical recovery was 102%. This method requires only 150 microliters of sample for both iron and TIBC which is a significant advantage over many existing methods in terms of sample volume, especially when a pediatric population is considered. Both iron and TIBC may be analyzed at the same time on the same rotor. The approximate analysis time is 30 minutes for seven iron and TIBC samples, four controls and one standard. The cost is low, being approximately $0.08 per iron assay and $0.83 per TIBC assay.

Properties and possible functions of the adenylate cyclase in plasma membranes of Saccharomyces cerevisiae.

We have examined the possible role of adenosine 3',5'-phosphate (cAMP) in functions associated with the plasma membranes of Saccharomyces cerevisiae. Purified membranes from this source contained an adenylate cyclase which was insensitive to activation by fluoride or guanine nucleotides, only weakly responsive to changes of carbon source in the growth medium, and strongly stimulated by vanadate. They also contained at least two classes of receptor proteins for guanine nucleotides (as measured by binding of labeled 5'-guanylyl methylene diphosphate) with apparent dissociation constants equal to 1.0 x 10(-7) and 3 x 10(-6) M, a protein kinase capable of phosphorylating added histones, the activity of which was stimulated by cAMP, and cAMP receptors that may function as regulatory subunits for this kinase. Membrane proteins were also susceptible to phosphorylation by endogenous kinase(s), with polypeptides of apparent molecular weights equal to 160 x 10(3), 135 x 10(3), 114 x 10(3), and 58 x 10(3) as the major targets. Of these, the 114,000-molecular-weight polypeptide was probably identical to the proton-translocating ATPase of the membranes. However, the cAMP-dependent protein kinase did not appear to be involved in these reactions. Intact (rho+ or rho0) cells responded to dissipation of the proton electrochemical gradient across their plasma membranes by rapid and transient changes in their intracellular level of cAMP, as suggested earlier (J. M. Trevillyan and M. L. Pall, J. Bacteriol., 138:397-403, 1979). Thus, although yeast plasma membranes contain all the essential components of a stimulus-responsive adenylate cyclase system, the precise nature of the coupling device and the targets involved remain to be established.

An enzymic, reaction-rate assay for serum creatinine with a centrifugal analyzer.

We describe a procedure for specific, rapid, kinetic determination of creatinine, in which a manual coupled-enzyme micro-scale assay is adapted to a centrifugal analyzer. The creatinine reaction is ultimately linked to NADH utilization, which is measured by the absorbance change at 340 nm. This procedure requires 15 microL of serum and the standard curve is linear to a creatinine concentration of 200 mg/L. A four-point kinetic algorithm allows the dynamic range of the assay to be extended without sacrificing sensitivity, and makes a separate serum blank unnecessary. The within-run precision (CV) for samples with a creatinine concentration of 11 and 52 mg/L was 5.6 and 2.4%, respectively; day-to-day CV for a creatinine concentration of 11 mg/L was 7.7% (n = 21). We compared this procedure with a kinetic Jaffé procedure, with excellent agreement (r = 0.996; y = 0.96x + 2.4 mg/L). Bilirubin, non-esterified fatty acids, and ketone bodies do not affect creatinine determinations by this method; thus the method is especially useful for monitoring the renal function of diabetics.

Partial characterization of the plasma membrane ATPase from a rho0 petite strain of Saccharomyces cerevisiae.

Crude membrane preparations of a rho0 mutant of Saccharomyces cerevisiae exhibit Mg2+-dependent ATPase activity. Over the optimal pH range, 5.0-6.75, the apparent Vmax of the enzyme equals 590 nmoles of ATP hydrolyzed per minute per milligram protein, with an apparent Km for ATP of 1.3 mM. ATP hydrolysis is insensitive to ouabain, venturicidin, aurovertin, and the protein inhibitor described by Pullman and Monroy; inhibited by oligomycin (at high concentrations) and sodium orthovanadate, and it is sensitive to dicyclohexylcarbodiimide, p-hydroxymercuribenzoate, hydroxylamine, sodium fluoride, and sodium iodoacetate. The pH optimum and the inhibitor pattern distinguish the plasma membrane enzyme from the mitochondrial F1 ATPase still present in these cells (this activity is sensitive to efrapeptin, aurovertin, and the protein inhibitor, but resistant to DCCD). In addition, the activity of the plasma membrane enzyme and its affinity for ATP are responsive to changes in the composition of the growth medium, with the highest activity observed in cells grown on methyl-alpha-D-glucoside, a sugar which results not only in partial release from catabolite repression but also requires the induction of an active transport system for growth.