Preferential exclusion of sucrose from recombinant interleukin-1 receptor antagonist: role in restricted conformational mobility and compaction of native state

Understanding the mechanism for sucrose-induced protein stabilization is important in many diverse fields, ranging from biochemistry and environmental physiology to pharmaceutical science. Timasheff and Lee [Lee, J. C. & Timasheff, S. N. (1981) J. Biol. Chem. 256, 7193-7201] have established that thermodynamic stabilization of proteins by sucrose is due to preferential exclusion of the sugar from the protein's surface, which increases protein chemical potential. The current study measures the preferential exclusion of 1 M sucrose from a protein drug, recombinant interleukin 1 receptor antagonist (rhIL-1ra). It is proposed that the degree of preferential exclusion and increase in chemical potential are directly proportional to the protein surface area and that, hence, the system will favor the protein state with the smallest surface area. This mechanism explains the observed sucrose-induced restriction of rhIL-1ra conformational fluctuations, which were studied by hydrogen-deuterium exchange and cysteine reactivity measurements. Furthermore, infrared spectroscopy of rhlL-1ra suggested that a more ordered native conformation is induced by sucrose. Electron paramagnetic resonance spectroscopy demonstrated that in the presence of sucrose, spin-labeled cysteine 116 becomes more buried in the protein's interior and that the hydrodynamic diameter of the protein is reduced. The preferential exclusion of sucrose from the protein and the resulting shift in the equilibrium between protein states toward the most compact conformation account for sucrose-induced effects on rhIL-1ra.     

Planar-flow spin-casting of molten metals: process behaviour

An experimental study of the planar-flow melt-spinning process was performed in order to gain a better understanding of the steady-state production of microcrystalline and amorphous ribbons. The dependence of the thickness of the ribbon product,T, on process parameters (wheel speed,U, nozzle/wheel gap,G, overpressure, ΔP, nozzle-slot breadth,R, and nozzle-slot width,W) was determined using an apparatus designed to deliver reproducible results. Thicknesses were reproducible to within 5%–8%. Guided by dimensional analysis, the non-dimensional thickness (T/G) was found to depend, within the experimental error, only on a non-dimensional pressure drop (ΔP/ρU ²) and slot breadth (R/G) for fixed thermal conditions. Data from the literature and our data, which considerably extend the range, correlate consistently on this basis. In contrast to the steady behaviour, the limits within which a uniform ribbon can be formed depend on a larger set of parameters; this dependence is sketched with the available data. Finally, a variety of observed ribbon surface textures (free meniscus side) is catalogued.     

A circuit approach to field computation

Many design objectives can be expressed more directly in terms of circuit parameters, such as capacitance and inductance, than in terms of the field vectors E and B. A simple, although radical, change is suggested to provide a closer link between the field and circuit views. The electric potential φ and magnetic vector A are defined, in energy terms, as the primary field quantities, and they are visualized as measures of the capacitance and inductance, respectively. E and B become auxiliary symbols denoting derivatives, so that the customary roles of the potentials and field vectors are reversed. Some of the practical advantages of the change are examined and are illustrated by example     

ART-EMAP: A neural network architecture for object recognition byevidence accumulation

A new neural network architecture is introduced for the recognition of pattern classes after supervised and unsupervised learning. Applications include spatio-temporal image understanding and prediction and 3D object recognition from a series of ambiguous 2D views. The architecture, called ART-EMAP, achieves a synthesis of adaptive resonance theory (ART) and spatial and temporal evidence integration for dynamic predictive mapping (EMAP). ART-EMAP extends the capabilities of fuzzy ARTMAP in four incremental stages. Stage 1 introduces distributed pattern representation at a view category field. Stage 2 adds a decision criterion to the mapping between view and object categories, delaying identification of ambiguous objects when faced with a low confidence prediction. Stage 3 augments the system with a field where evidence accumulates in medium-term memory. Stage 4 adds an unsupervised learning process to fine-tune performance after the limited initial period of supervised network training. Each ART-EMAP stage is illustrated with a benchmark simulation example, using both noisy and noise-free data.     

Transient conduction in a slab with chemical reaction

The heat-balance integral technique is used to analyse a one-dimensional transient conduction process. Heat is transferred by convection to the surface of a slab and when the surface temperature reaches a certain value, an endothermic reaction is triggered which propagates as an isothermal front in the same direction as the conduction front. Two possible routes to the fully developed heat transfer condition are treated in a way which allows the approximate temperature distribution in the slab to be predicted as a function of the main process variables.     

Analytical electron microscopy of graphite-rich inclusions in sintered α-silicon carbide

Intragranular inclusions and multiphase regions at triple grain junctions in (B+C)-doped sintered -SiC were investigated using analytical and high-resolution electron microscopy. Both regions were two-phase, composed of graphite and amorphous material. The triple junctions also contained pores. The amorphous regions were principally carbon and oxygen. Graphite was formed by partial transformation of the amorphous regions. Only the triple-junction regions contain typical impurities from the starting -SiC powder, inferring that they are the main sinks for all grain-boundary/surface impurities in the material system.     

Effect of added water, sodium erythorbate and storage time on the functional properties of prerigor beef preblends in a model system

Singular and combined effects of added water, sodium erythorbate and storage time on salt soluble protein extractability, bacteriological and chemical characteristics of preblended hot-boned beef were evaluated. Waterholding and gel forming capacities of preblended hot-boned beef containing either 0, 10 or 20% added water were determined. Significant interactions between added water level and storage time on microbial counts and between sodium erythorbate level and storage time on thiobarbituric acid values and residual nitrite levels were noted. The presence of sodium erythorbate resulted in a more (P < 0·05) rapid rate of nitrite disappearance, but it did not affect (P > 0·05) microbial counts. Salt-soluble protein extractability was not affected (P > 0·05) by added water, but more protein could be extracted with increasing storage time. A trend existed to suggest that the presence of added water in the meat preblends slightly improved the gel formation and waterholding capacities.     

Tenderness of major muscles from three breed-types of cattle at different times-on-feed

Seventy-five steers (9 to 12 months of age) of Angus (n = 25), Brahman (n = 25) and Brahman × Angus (n = 25) breed-types of known history were fed a high-energy diet and five steers from each breed-type were slaughtered after 0, 56, 112, 168 and 224 days on test. At seven days post mortem, the left side of each carcass was fabricated and eight major primals or subprimals were obtained. Steaks were removed from the shoulder clod, strip loin, tenderloin, top sirloin, knuckle, top round, bottom round and eye of round for Warner-Bratzler shear (WBS) determinations and from the strip loin and bottom round for sensory panel evaluations. Steaks from Angus and Brahman × Angus steers were generally more tender than steaks from Brahman steers. Tenderness of steaks from steers of the three breed-types responded to time-on-feed differently: (a) steaks from Brahman steers improved (P < 0·05) in tenderness with increasing time-on-feed for all muscles except the M. psoas major (tenderloin); (b) with the exception of the M. longissimus muscle (strip loin), the tenderness of steaks from Angus steers did not change (P > 0·05) as time-on-feed increased and (c) for the Brahman × Angus steers, five of the ten muscles studied improved (P < 0·05) in tenderness with increasing time-on-feed. 'Acceptable' tenderness (WBS values of less than 4·5 kg) for most of the muscles was achieved at feeding periods from 112 to 224 days for Brahman, from 0 to 56 days for Angus and from 56 to 168 days for Brahman × Angus.     

Carcass characteristics and composition of Brahman, angus and Brahman x Angus steers fed for different times-on-feed

Twenty-five steers of each of three breedtypes (Angus, Brahman and F(1) Brahman x Angus) were sorted by frame size and muscle thickness, assigned to groups (five steers of each breedtype) to be fed for 0, 56, 112, 168 or 224 days, slaughtered and compared for various carcass traits. Steers of each breedtype had similar dressing percentages. Carcasses from all three breedtypes merited similar USDA quality and yield grades; breedtypes differences in quality grade were slight. Differences were found in the fat deposition patterns exhibited by the three breedtypes. Brahman steers tended to deposit more of their total fat as subcutaneous fat early in the feeding period. Angus steers had more (P < 0·05) seam fat as a percentage of carcass weight at all five feeding periods and more (P < 0·05) kidney, pelvic and heart fat at two of the five feeding periods than Brahman steers. Brahman steers had a higher percentage of their separable lean in the muscles of the round than did steers of the other breedtypes.