Abstract: Ultrafast, laser-induced pH jump with time-resolved photoacoustic detection has been used to investigate the early protonation steps leading to the formation of the compact acid intermediate (I) of apomyoglobin (ApoMb). When ApoMb is in its native state (N) at pH 7.0, rapid acidification induced by a laser pulse leads to two parallel protonation processes. One reaction can be attributed to the binding of protons to the imidazole rings of His24 and His119. Reaction with imidazole leads to an unusually large contraction of -82 +/- 3 ml/mol, an enthalpy change of 8 +/- 1 kcal/mol, and an apparent bimolecular rate constant of (0.77 +/- 0.03) x 10(10) M(-1) s(-1). Our experiments evidence a rate-limiting step for this process at high ApoMb concentrations, characterized by a value of (0. 60 +/- 0.07) x 10(6) s(-1). The second protonation reaction at pH 7. 0 can be attributed to neutralization of carboxylate groups and is accompanied by an apparent expansion of 3.4 +/- 0.2 ml/mol, occurring with an apparent bimolecular rate constant of (1.25 +/- 0.02) x 10(11) M(-1) s(-1), and a reaction enthalpy of about 2 kcal/mol. The activation energy for the processes associated with the protonation of His24 and His119 is 16.2 +/- 0.9 kcal/mol, whereas that for the neutralization of carboxylates is 9.2 +/- 0.9 kcal/mol. At pH 4.5 ApoMb is in a partially unfolded state (I) and rapid acidification experiments evidence only the process assigned to carboxylate protonation. The unusually large contraction and the high energetic barrier observed at pH 7.0 for the protonation of the His residues suggests that the formation of the compact acid intermediate involves a rate-limiting step after protonation.

Abstract: We have developed an instrumental setup that uses transient absorption to monitor protein folding/unfolding processes following a laser-induced, ultrafast release of protons from o-nitrobenzaldehyde. The resulting increase in [H(+)], which can be more than 100 microM, is complete within a few nanoseconds. The increase in [H(+)] lowers the pH of the solution from neutrality to approximately 4 at the highest laser pulse energy used. Protein structural rearrangements can be followed by transient absorption, with kinetic monitoring over a broad time range (approximately 10 ns to 500 ms). Using this pH-jump/transient absorption technique, we have examined the dissociation kinetics of non-native axial heme ligands (either histidine His26 or His33) in GuHCl-unfolded Fe(III) cytochrome c (cyt c). Deligation of the non-native ligands following the acidic pH-jump occurs as a biexponential process with different pre-exponential factors. The pre-exponential factors markedly depend on the extent of the pH-jump, as expected from differences in the pK(a) values of His26 and His33. The two lifetimes were found to depend on temperature but were not functions of either the magnitude of the pH-jump or the pre-pulse pH of the solution. The activation energies of the deligation processes support the suggestion that GuHCl-unfolded cyt c structures with non-native histidine axial ligands represent kinetic traps in unfolding.

Abstract: This volume brings together studies from various disciplines of the social sciences and humanities (Anthropology, Sociology, Cultural Studies, History and Literary theory) that examine the equestrian world as a historically gendered and highly dynamic field of contemporary sport and culture. From elite international dressage and jumping, polo and the turf, to the rodeo world of the Americas and popular forms of equestrian sport and culture, we are introduced to a range of issues as they unfold at local and global, national and international levels. Students and scholars of gender, culture and sport will find much of interest in this original look at contemporary issues such as “engendered” (women’s and men’s) dentities/subjectivities of equestrians, representations of girls, horses and the world of adventure in juvenile fiction; the current “feminization” of particular equestrian activities (and where boys and men stand in relation to this); how broad forms of social inequality and stratification play themselves out within gendered equestrian contexts; men and women and their relation to horses within the framework of current discussions on the relation of animals to humans (which may include not only love and care, but also exploitation and violence), among others. Singular contributions that incorporate a wide variety of classic and contemporary theoretical perspectives and empirical methodologies show how horse cultures around the globe contribute to historical and current constructions of embodied “femininities” and “masculinities”, reflecting a world that has been moving “beyond the binaries” while continuing to be enmeshed in their persistent and contradictory legacy. The final chapter makes a brave attempt at synthesizing individual chapters and moving forward from the evidences they provide, to suggest a compelling agenda for future research.

Abstract: The ability of horses to learn through social observation may ease the implementation of new management systems, because the use of automatic feeders etc. by naive horses could be facilitated by observation of experienced horses. However, previous studies found no documentation for observational learning abilities in horses. This study aimed to investigate the ability of horses to learn an instrumental task from a familiar conspecific when social interaction was allowed during the demonstration. Two similar experiments were performed. In the first experiment, Observer horses (n=11) participated in ten successive demonstrations, where a trained Demonstrator opened an operant device by pushing a sliding lid aside with the muzzle in order to obtain a food reward. Immediately after the demonstrations the Observer horses were given the opportunity to operate the device alone. Control horses (n=11) were aware that the device contained food but were presented to the operant device without demonstration of the task. The learning criterion was at least two openings. Accomplishment of and latency to accomplish the learning criterion, and investigative behaviour towards the operant device were recorded. Five Observers and one Control, out of the eleven horses in each treatment group, accomplished the learning criterion. Even though this presents a high odds ratio (OR) in favour of the Observer treatment (OR=7.6), there was no significant difference between the treatment groups (P=0.15). Analysis of investigative behaviour showed, however, that the demonstrations increased the motivation of the Observer horses to investigate the device. Subsequently, a similar experiment was performed in a practical setting with 44 test horses (mixed age, gender and breed). We used the same operant device and the same number and type of demonstrations, although the horses were held on a loose rope to minimise aggression. In this second experiment, six of 23 Observer horses and five of 21 Control horses learned the instrumental task, representing no influence of the demonstration. Thus, this study did not demonstrate an ability of horses to learn an instrumental task through observation.

Abstract: Abstract Concentrated feed diets have been shown to drastically increase the rate of the cribbing, an oral stereotypy in horses, but the specific component causing the rise has not been identified. Furthermore, the mechanism through which feed affects cribbing has not been explored. In the first experiment of this study, we quantified the latency to crib and number of cribs in 15 min after the horses tasted various grain, sugar, and artificial sweetener solutions. Undiluted grain stimulated the most cribs (P < 0.01) compared with all other solutions, and shortest latency to crib, although this was significantly higher only when compared with diluted grain (P = 0.03). In Experiment 2, latency to crib and number of cribs in 15 min after the grain and sugar solutions were administered via nasograstric tube were also evaluated. There were no statistical differences among cribbing responses to grain, fructose, and water administered directly to the stomach although grain stimulated cribbing behavior more quickly than 10% fructose (P = 0.03) and 100% tap water (P = 0.04). These results confirm that highly palatable diets, possibly mediated through the opioid and dopaminergic systems, are one of the most potent inducers of cribbing behavior. The highly palatable taste remains the probable “cribogenic” factor of concentrated diet, although gastric and post-gastric effects cannot be excluded.