Isolation, physicochemical properties, and the macromolecular composition of the vitelline and fertilization envelopes from Xenopus laevis eggs

As a step toward defining in molecular terms the sperm-triggered block to polyspermy reaction established by the egg at fertilization, vitelline (VE) and fertilization (FE) envelopes were isolated from eggs of the Sounth African clawed toad Xenopus laevis and some of their physicochemical properties determined. Envelopes were isolated after lysis of the fertilized or unfertilized eggs by sieving techniques; isolated envelopes retained their in situ morphology as determined by electron microscopy. The isolated envelopes had different solubility properties and, in general, VE was more readily dissolved by aqueous solvents than FE, although both could be completely dissolved by detergents or chaotropic agents. Changes in envelope solubility correlated with the progression of the cortical reaction implicating a role for cortical granule material in modifying the solubility properties of the envelope. The VE and FE were composed of protein and carbohydrate with no lipid components detected. As determined by immunodiffusion experiments, the FE contained the same antigens as the VE plus components derived from the cortical granules and the innermost jelly layer, J. The macromolecular composition of the envelopes was determined by sodium dodecyl sulfate gel electrophoresis. The VE contained at least 11 glycoproteins with molecular weights ranging from 125 000 to less than 16 000 with two components (40 000 and 33 000) accounting for almost two-thirds of the total stainable material. The FE contained ten glycoproteins that had the same molecular weights as those in the VE. One glycoprotein component underwent a reduction in molecular weight from 77 000 to 67 500 when the VE was converted to the FE. This molecular weight change was interpreted as the probable result of limited proteolysis. In addition, the FE gel electrophoresis patterns contained macromolecular components derived from the cortical granules and jelly layer, J, consistent with the immunodiffusion experiments. These components were absent when the FE was prepared in the absence of Ca2+, suggesting a role for Ca2+ in binding the VE, cortical granules, and J components together. We concluded that the conversion of the glycoproteinaceous VE to FE at fertilization is caused by interaction of the VE with components from the cortical granules and jelly layer J. These interactions are of both a chemical and physical nature.     

Feeding rolled barley grain steeped in lactic acid modulated energy status and innate immunity in dairy cows

Feeding dairy cows large proportions of cereal grain is commonly associated with rumen acidosis, activation of innate immunity, and perturbation of intermediary metabolism. We previously showed that steeping barley grain in 0.5% lactic acid (LA) decreased the rate of starch degradation, lowered the risk of subacute rumen acidosis, modulated rumen fermentation profile, and increased milk fat content in dairy cows. This study sought to investigate whether feeding of LA-treated barley grain would affect carbohydrate and lipid metabolism as well as innate immunity. Eight rumen-fistulated late-lactation (approximately 217 d in milk, DIM) Holstein cows were randomly assigned, in a 2 × 2 crossover design, to 1 of the 2 dietary treatments consisting of 27% (dry matter basis) rolled barley grain steeped for 48 h in an equal volume (wt/vol) of tap water (CTR) or 0.5% LA (TRT). Each experimental period lasted 21 d, with the first 11 d for diet adaptation. Blood and rumen samples were collected on d 12, 15, 17, and 21 of the experimental period before the morning feeding to evaluate the effects of dietary treatment on preprandial day-to-day variation of plasma and rumen variables. To establish the effect of treatment on diurnal variation of plasma variables, blood samples were collected on the last day of each period at 0, 2, 4, 6, 8, 10, and 12 h after the morning feeding (i.e., 0800 h). Results of the day-to-day study showed that cows fed the TRT diet had greater overall preprandial concentrations of glucose, cholesterol, and insulin, and a lower concentration of haptoglobin in plasma. Diurnal data indicated lower concentrations of haptoglobin and serum amyloid A and a tendency for greater plasma lactate in cows fed the TRT diet. A treatment by time interaction was observed for glucose, lactate, insulin, haptoglobin, and lipopolysaccharide-binding protein, suggesting a role for both the processing of grain and the time of sampling on those variables. No effect of diet on plasma concentrations of cortisol, β-hydroxybutyrate, and nonesterified fatty acids or rumen endotoxin was evidenced. Taken together, our results demonstrated that feeding barley grain steeped in 0.5% LA modulated both energy status and innate immunity of dairy cows fed relatively high levels (45% of dry matter) of dietary concentrate.     

β-Galactosidase activity of commercial lactase samples in raw and pasteurized milk at refrigerated temperatures

Many consumers are unable to enjoy the benefits of milk due to lactose intolerance. Lactose-free milk is available but at about 2 times the cost of regular milk or greater, it may be difficult for consumers to afford. The high cost of lactose-free milk is due in part to the added cost of the lactose hydrolysis process. Hydrolysis at refrigerated temperatures, possibly in the bulk tank or package, could increase the flexibility of the process and potentially reduce the cost. A rapid β-galactosidase assay was used to determine the relative activity of commercially available lactase samples at different temperatures. Four enzymes exhibited low-temperature activity and were added to refrigerated raw and pasteurized milk at various concentrations and allowed to react for various lengths of time. The degree of lactose hydrolysis by each of the enzymes as a function of time and enzyme concentration was determined by HPLC. The 2 most active enzymes, as determined by the β-galactosidase assay, hydrolyzed over 98% of the lactose in 24 h at 2°C using the supplier's recommended dosage. The other 2 enzymes hydrolyzed over 95% of the lactose in 24 h at twice the supplier's recommended dosage at 2°C. Results were consistent in all milk types tested. The results show that it is feasible to hydrolyze lactose during refrigerated storage of milk using currently available enzymes.     

Emotional reactions to surgery.

Evaluated the stress effects of major surgery on 26 white male patients. The State-Trait Anxiety Inventory (STAI) and Kincannon's Mini-Mult version of the MMPI were given 18-24 hr. before surgery and 3-9 days postsurgery after the S was informed he was recovering without complications. Mean STAI A-State scores were much higher prior to surgery than after; STAI A-Trait scores were essentially the same. Patients with high- and low-A-Trait scores showed similar presurgery-postsurgery changes in A-State. Scores on the Mini-Mult were essentially unchanged by the stresses associated with surgery. Results indicate that the threat of imminent surgery produced elevations in anxiety as an emotional state, but did not affect anxiety proneness (A-Trait). (27 ref.) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Electromechanical Properties of Porous Piezoelectric Ceramics

A theoretical approach is forwarded to predict the electromechanical properties of porous piezoelectric ceramics. The analysis is able to account for the effects of porosity shape and concentration and is applicable to piezoelectric ceramics of arbitrary material symmetry. By coupling the exact solution for a single ellipsoidal pore embedded in an infinite piezoelectric matrix with an effective medium approximation, the theory considers, in an approximate manner, interaction effects at finite porosity concentrations. The theoretical estimates are developed using a matrix formulation which enables all elastic, dielectric, and piezoelectric moduli of the porous solid to be readily computed. Numerical results are presented to illustrate the effects of the shape and concentration of the porosity on the effective electroelastic moduli and transducer parameters of practical importance. Particular attention is devoted to assessing the sensitivity of the effective electromechanical properties to the accuracy of the input data. Finally, theoretical estimates are shown to be in good agreement with existing experimental results for porous piezoelectric ceramics with various microstructural geometries.     

Structural studies on an inhibitory antibody against Thermus aquaticus DNA polymerase suggest mode of inhibition

TP7, an antibody against Thermus aquaticus DNA polymerase I (TaqP), is usedas a thermolabile switch in 'hot start' variations of PCR to minimizenon-specific amplification events. Earlier studies have established thatTP7 binds to the polymerase domain of TaqP, competes with primer templatecomplex for binding and is a potent inhibitor of the polymerase activity ofTaqP. We report crystallographic determination of the structure of an Fabfragment of TP7 and computational docking of the structure with the knownthree-dimensional structure of the enzyme. Our observations stronglysuggest that the origin of inhibitory ability of TP7 is its binding toenzyme residues involved in DNA binding and polymerization mechanism.Although criteria unbiased by extant biochemical data have been used inidentification of a putative solution, the resulting complex offers aneminently plausible structural explanation of biochemical observations. Theresults presented are of general significance for interpretation of dockingexperiments and in design of small molecular inhibitors of TaqP, that arenot structurally similar to substrates, for use in PCR. Structural andfunctional similarities noted among DNA polymerases, and the fact thatseveral DNA polymerases are pharmacological targets, make discovery ofnon-substrate based inhibitors of additional importance.     

Interfacial behaviour of various liquid electrode/β″-alumina systems

Three different liquid electrode/sodium-alumina interfaces were investigated. The NaNO₃/-alumina interface displays ohmic behaviour although the resistance is somewhat larger than expected. The sodium/-alumina interface exhibits asymmetric resistance with respect to d.c. current flow. Cyclic voltammetry results suggest that a Na₂O film is being modified electrochemically during the sweeps. When a sodium amalgam electrode is utilized, the film, if it is present, has very little effect. The asymmetric resistance is not observed and there is good agreement between d.c. and a.c. resistances after only a small amount of sodium deposition.     

Direct calculation of the sulphur electrode resistance in sodium-sulphur cells

A simple model is used to compute the resistance of the sulphur electrode as a function of charge. It is shown that the general network used for the simulation of the behaviour of the sulphur electrode can be simplified because of the relatively small value of the polarization resistance. Reasonable assumptions concerning the distribution of reactants and products are discussed. The model is able to assess the contribution of the sulphur electrode to the total cell resistance for a 16 Ah capacity cell.