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Abbruzzetti, S., Crema, E., Masino, L., Vecli, A., Viappiani, C., Small, J. R., et al. (2000). Fast events in protein folding: structural volume changes accompanying the early events in the N-->I transition of apomyoglobin induced by ultrafast pH jump. Biophys J, 78(1), 405–415.
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.
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Abbruzzetti, S., Viappiani, C., Sinibaldi, F., & Santucci, R. (2004). Kinetics of histidine dissociation from the heme Fe(III) in N-fragment (residues 1-56) of cytochrome c. Protein J, 23(8), 519–527.
Abstract: We have here investigated the dissociation kinetics of the His side chains axially ligated to the heme-iron in the ferric (1-56 residues) N-fragment of horse cyt c. The ligand deligation induced by acidic pH-jump occurs as a biexponential process with different pre-exponential factors, consistent with a structural heterogeneity in solution and the presence of two differently coordinated species. In analogy with GuHCl-denatured cyt c, our data indicate the presence in solution of two ferric forms of the N-fragment characterized by bis-His coordination, as summarized in the following scheme: His18-Fe(III)-His26 <==> His18-Fe(III)-His33. We have found that the pre-exponential factors depend on the extent of the pH-jump. This may be correlated with the different pKa values shown by His26 and His33; due to steric factors, His26 binds to the heme-Fe(III) less strongly than His33, as recently shown by studies on denatured cyt c. Interestingly, the two lifetimes are affected by temperature but not by the extent of the pH-jump. The lower pKa for the deligation reaction required the use of an improved laser pH-jump setup, capable of inducing changes in H+ concentration as large as 1 mM after the end of the laser pulse. For the ferric N-fragment, close activation entropy values have been determined for the two histidines coordinated to the iron; this result significantly differs from that for GuHCl-denatured cyt c, where largely different values of activation entropy were calculated. This underlines the role played by the missing segment (residues 57-104) peptide chain in discriminating deligation of the “nonnative” His from the sixth coordination position of the metal.
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Abbruzzetti, S., Viappiani, C., Small, J. R., Libertini, L. J., & Small, E. W. (2001). Kinetics of histidine deligation from the heme in GuHCl-unfolded Fe(III) cytochrome C studied by a laser-induced pH-jump technique. J Am Chem Soc, 123(27), 6649–6653.
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.
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Aberle, K. S., Hamann, H., Drögemüller, C., & Distl, O. (2004). Genetic diversity in German draught horse breeds compared with a group of primitive, riding and wild horses by means of microsatellite DNA markers. Anim. Gen., 35(4), 270–277.
Abstract: Summary We compared the genetic diversity and distance among six German draught horse breeds to wild (Przewalski's Horse), primitive (Icelandic Horse, Sorraia Horse, Exmoor Pony) or riding horse breeds (Hanoverian Warmblood, Arabian) by means of genotypic information from 30 microsatellite loci. The draught horse breeds included the South German Coldblood, Rhenish German Draught Horse, Mecklenburg Coldblood, Saxon Thuringa Coldblood, Black Forest Horse and Schleswig Draught Horse. Despite large differences in population sizes, the average observed heterozygosity (Ho) differed little among the heavy horse breeds (0.64�0.71), but was considerably lower than in the Hanoverian Warmblood or Icelandic Horse population. The mean number of alleles (NA) decreased more markedly with declining population sizes of German draught horse breeds (5.2�6.3) but did not reach the values of Hanoverian Warmblood (NA = 6.7). The coefficient of differentiation among the heavy horse breeds showed 11.6% of the diversity between the heavy horse breeds, as opposed to 21.2% between the other horse populations. The differentiation test revealed highly significant genetic differences among all draught horse breeds except the Mecklenburg and Saxon Thuringa Coldbloods. The Schleswig Draught Horse was the most distinct draught horse breed. In conclusion, the study demonstrated a clear distinction among the German draught horse breeds and even among breeds with a very short history of divergence like Rhenish German Draught Horse and its East German subpopulations Mecklenburg and Saxon Thuringa Coldblood.
Keywords: diversity; endangered breeds; genetic variation; horse; microsatellite
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Abeyesinghe, S. M., Nicol, C. J., Hartnell, S. J., & Wathes, C. M. (2005). Can domestic fowl, Gallus gallus domesticus, show self-control? Anim. Behav., 70(1), 1–11.
Abstract: An important aspect of cognition is whether animals live exclusively in the present or can anticipate the future. Defined as self-control, the ability to choose a large, remote reinforcer over a small, proximate reinforcer available at the same frequency has been examined in a number of species, often proving difficult to demonstrate. We investigated self-control for food in domestic fowl using a standard two-key operant task and an equivalent two-choice return maze (TCRM) task. When hens chose between a 2-s delay to a 3-s feed access (impulsive) and a 6-s delay to a 7-s feed access (self-control), they appeared unable to discriminate in the TCRM but were impulsive in the operant task. We explored reasons for not choosing self-control in the operant task, first by examining the relation between feed access time and actual feed intake. A second operant experiment examined whether failure to show self-control could be attributed to an inability to combine the delay and access (quantity) reward information associated with choices to reach overall predictions of value. New hens chose between a 2-s delay to a 3-s feed access (impulsive) and either a 22-s delay to a 22-s feed access (standard self-control) or a 6-s delay to a 22-s feed access (jackpot self-control). While hens were impulsive in the standard condition, they showed significant and pronounced self-control in the jackpot condition, eliminating the possibility of an absolute cognitive constraint. Impulsive behaviour can instead be explained by temporal discounting: perceived depreciation of reward value as a function of the uncertainty associated with delay. Implications for welfare are discussed.
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Abeyesinghe, S. M., Nicol, C. J., Wathes. C.M., & Randall, J. M. (2001). Development of a raceway method to assess aversion of domestic fowl to concurrent stressors. Behav. Process., 56(3), 175–194.
Abstract: The requirement for assessing the effects of stressor combinations in improving the welfare of animals has not been widely recognised. Knowledge of the effects of concurrent stressors is needed to improve environments such as transport, where animals are presented with many simultaneous challenges. However, no method for measuring the effects of different stressors with a common unit is currently available. A locomotor passive avoidance method was developed as a common currency measure of the aversion of domestic fowl to concurrent stressors, using vibrational and thermal stressors as an exemplar. Juvenile fowl, fasted overnight, were trained to run a raceway into a goal-box for small food rewards (FR1). When running consistently, the reinforcement schedule was superimposed with a FR5 treatment schedule (60 min confinement in the goal-box with either a control of no other stressors [N] or concurrent vibration and thermal stressors [VT]). Subsequent latency to return to the goal-box was recorded as a measure of aversion. The factors affecting bird response were addressed in a series of experiments to optimise the method and clarify interpretation of results. Pre-feeding (20% ration 2 h prior to testing) did not affect response, but increasing the number of treatment presentations facilitated learning and increased method sensitivity. Treatment responses were consistent across experiments; overall VT was avoided (P<0.001), but N was not. However, there was large individual variation in response to VT. A final experiment indicated that, given a visual discriminatory cue, birds were capable of learning the required association between entering the goal-box and receiving the treatment, suggesting that the delay responses were due to aversion rather than the immediate impact of treatment on ability to respond. Further work is required to test the singular stressors, but the method retains common currency potential for assessing aversion to multiple stressors.
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Adamo, S. A., Ehgoetz, K., Sangster, C., & Whitehorne, I. (2006). Signaling to the Enemy? Body Pattern Expression and Its Response to External Cues During Hunting in the Cuttlefish Sepia officinalis (Cephalopoda). Biol. Bull., 210(3), 192–200.
Abstract: Abstract. Cuttlefish can rapidly alter their appearance by using neurally controlled chromatophore organs. This ability may provide a window into their cognitive capacity. We test whether the changes in body pattern that occur during hunting depend on context. If they do, then it may be possible to use these changes to study cephalopod cognition while the animal is engaged in ecologically relevant tasks. We found consistent individual differences in the tendency of cuttlefish to hunt with the first two arms raised. We also found that cuttlefish usually darken their skin after they seize a prey item. This darkening is observed regardless of the identity of the prey (fish, crab, or shrimp), prey context (buried in sand, in a bare tank, or on top of a rock pile), or the presence of a sudden stimulus. The sudden stimulus was created by presenting an overhead model bird to the cuttlefish. The model induced components of the Deimatic Display, which is a form of antipredator behavior, suggesting that the model was perceived as a potential threat. Passing Cloud displays and the Darkening of the arms were significantly reduced after exposure to the model bird. The effect of a potential predator on body pattern expression during hunting suggests it may be possible to use these changes as a sensitive indicator of ecologically relevant learning. N1 -
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Agrillo, C., Dadda, M., & Bisazza, A. (2007). Quantity discrimination in female mosquitofish. Anim. Cogn., 10(1), 63–70.
Abstract: The ability in animals to count and represent different numbers of objects has received a great deal of attention in the past few decades. Cumulative evidence from comparative studies on number discriminations report obvious analogies among human babies, non-human primates and birds and are consistent with the hypothesis of two distinct and widespread mechanisms, one for counting small numbers (<4) precisely, and one for quantifying large numbers approximately. We investigated the ability to discriminate among different numerosities, in a distantly related species, the mosquitofish, by using the spontaneous choice of a gravid female to join large groups of females as protection from a sexually harassing male. In one experiment, we found that females were able to discriminate between two shoals with a 1:2 numerosity ratio (2 vs. 4, 4 vs. 8 and 8 vs. 16 fish) but failed to discriminate a 2:3 ratio (8 vs. 12 fish). In the second experiment, we studied the ability to discriminate between shoals that differed by one element; females were able to select the larger shoal when the paired numbers were 2 vs. 3 or 3 vs. 4 but not 4 vs. 5 or 5 vs. 6. Our study indicates that numerical abilities in fish are comparable with those of other non-verbal creatures studied; results are in agreement with the hypothesis of the existence of two distinct systems for quantity discrimination in vertebrates.
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Ahrendt, L. P., Labouriau, R., Malmkvist, J., Nicol, C. J., & Christensen, J. W. (2015). Development of a standard test to assess negative reinforcement learning in horses. Appl. Anim. Behav. Sci., 169, 38–42.
Abstract: Most horses are trained by negative reinforcement. Currently, however, no standardised test for evaluating horses' negative reinforcement learning ability is available. The aim of this study was to develop an objective test to investigate negative reinforcement learning in horses. Twenty-four Icelandic horses (3 years old) were included in this study. The horses were tested in a pressure-release task on three separate days with 10, 7 and 5 trials on each side, respectively. Each trial consisted of pressure being applied on the hindquarter with an algometer. The force of the pressure was increased until the horse moved laterally away from the point of pressure. There was a significant decrease in required force over trials on the first test day (P<0.001), but not the second and third day. The intercepts on days 2 and 3 differed significantly from day 1 (P<0.001), but not each other. Significantly stronger force was required on the right side compared to the left (P<0.001), but there was no difference between first and second side tested (P=0.56). Individual performance was evaluated by median-force and the change in force over trials on the first test day. These two measures may explain different characteristics of negative reinforcement learning. In conclusion, this study presents a novel, standardised test for evaluating negative reinforcement learning ability in horses.
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Ajie, B. C., Pintor, L. M., Watters, J., Kerby, J. L., Hammond, J. I., & Sih, A. (2007). A framework for determining the fitness consequences of antipredator behavior. Behav. Ecol., 18(1), 267–270.
Abstract: Behavioral ecologists have long been interested in understanding the adaptive value of antipredator behavior (Sih 1987Go; Lima and Dill 1990Go; Lima 1998Go). A recent review by Lind and Cresswell (2005)Go, however, noted some important difficulties with quantifying the fitness consequences of antipredator behaviors. In essence, Lind and Cresswell suggest that most studies do not provide strong evidence on the adaptive value of antipredator behavior because they do not consider 1) trade-offs between antipredator and reproductive performance, 2) the abilities of organisms to avoid fitness losses associated with constraints on focal traits by employing behavioral alternatives (behavioral compensation), and 3) the effects of behavioral defenses at different stages of the predation sequence. The authors rightfully assert that an understanding of these issues can only be accomplished by measuring multiple traits and fitness components (i.e., survival and reproduction). Nevertheless, the question of how to integrate such data into
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