Abstract: The acidic transition of porcine plasminogen, prepared by affinity chromatography, was studied by non-destructive methods. These methods are based on the analysis of the behaviour of the tryptophyls under various conditions. The perturbation of the absorption and emission spectra by pH or temperature and the dynamic quenching of the intrinsic fluorescence are used to obtain information on structural changes which affect the environment of these residues. It is shown that by decreasing pH the fluorescence emission spectra are shifted toward the long wavelengths, with a broadening of the fluorescence band. The same effect can be obtained at constant pH by heating the protein solution. In order to analyze these phenomena, it is assumed that the fluorescence intensities at 355 nm and 328 nm reflect the proportion of the tryptophans which are exposed to the solvent, and buried, respectively. The plot of the ratio of the fluorescence intensities at these wavelengths versus pH or temperature leads to a titration curve showing an unmasking of tryptophans. The proportion of exposed tryptophans is measured by the dynamic fluorescence quenching technique and the data analyzed according to Lehrer. The plot of the fraction of exposed tryptophyls versus pH also shows the unmasking of these chromophores. Thermal perturbation of a solution of plaminogen at neutral pH induces a difference absorption spectrum whose amplitudes at the maxima are proportional to the number of exposed aromatic residues. The comparison with a solution of fully denatured plasminogen in 6 M guanidium chloride, where all the tryptophyls are exposed, shows that the percentage of exposure is equal to 59%. This number is significantly higher than the percentage found by the fluorescence quenching technique (20%), indicating that some tryptophyls are located in crevices, exposed to the solvent but not to the iodide. At acidic pH the absorption difference spectra induced by thermal perturbation are not classical, since they show an inversion and a new band between 300 nm and 305 nm. This band is mentioned in the literature as a minor band of tryptophan which appears when this chromophore is located in an asymmetric environment. On plotting the maximum amplitude of these spectra obtained at acidic pH versus temperature, we obtain a curve indicating that two types of antagonistic interactions are involved in the perturbation of the chromophores spectra. The spectrophotometric titration of plasminogen gives classical absorption difference spectra. By plotting the maximum amplitude at 292 nm versus pH, we obtain a titration curve with an apparent pK of 2.9 units. This pK is acidic which respect to the pK value of a normal carboxyl. This low value can be due to a positively charged group in the neighbourhood of a carboxyl, which interacts with one or more chromophores. When the carboxyl becomes protonated, this positively charged group is free and available to perturb the environment of some chromophores...

Abstract: The dangers of hypothesizing about unobservable cognitive mechanisms are well known to behavior analysts. I propose, however, that carefully fashioned cognitive theories that make predictions that are inconsistent with current behavioral theories can provide useful research tools for the understanding of behavior. Furthermore, even if the results of such research may be accommodated by modifying existing behavioral theories, our understanding of behavior is often advanced by the empirical findings because it is unlikely that the research would have been conducted in the absence of such cognitive hypothesizing. Two examples of the development of emergent relations are described: The first deals with the nature of a pigeon's 'representation' of two stimuli both of which are associated with correct responding to a third in a many-to-one matching task (stimulus equivalence or common representations). The second has to do with transitive inference, the emergent relation between two stimuli mediated by their relation to a common stimulus in a simultaneous discrimination.

Abstract: Six pregnant mares were used to determine what level of water restriction causes physiological and/or behavioural changes indicative of stress. Nonlegume hay was fed ad libitum. During the first week of restriction, 5 l water/100 kg bwt was available, during the second week 4 l/100 kg bwt and, during the third week, 3 l/100 kg bwt. Ad libitum water intake was 6.9 l/100 kg bwt; at 3 l/100 kg bwt water intake was 42% of this. Daily hay intake fell significantly with increasing water restriction from 12.9 +/- 0.75 kg to 8.3 +/- 0.54 kg; bodyweight fell significantly for a total loss of 48.5 +/- 8.3 kg in 3 weeks. Daily blood samples were analysed; osmolality rose significantly with increasing water restriction from 282 +/- 0.7 mosmols/kg to 293.3 +/- 0.8 mosmols/kg bwt, but plasma protein and PCV did not change significantly. Cortisol concentrations fell from 8.1 ng/ml to 6.4 ng/ml over the 3 week period. Aldosterone fell from 211.3 +/- 74.2 pg/ml to 92.5 +/- 27.5 pg/ml at the end of the first week. The behaviour of 4 of the 6 mares was recorded 24 h/day for the duration of the study. The only significant difference was in time spent eating, which decreased with increasing water restriction from 46 +/- 3% to 30 +/- 3%. It is concluded that water restriction to 4 l/100 kg bwt dehydrates pregnant mares and may diminish their welfare, but is not life- or pregnancy-threatening.