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Gulotta, M., Gilmanshin, R., Buscher, T. C., Callender, R. H., & Dyer, R. B. (2001). Core formation in apomyoglobin: probing the upper reaches of the folding energy landscape. Biochemistry, 40(17), 5137–5143.
Abstract: An acid-destabilized form of apomyoglobin, the so-called E state, consists of a set of heterogeneous structures that are all characterized by a stable hydrophobic core composed of 30-40 residues at the intersection of the A, G, and H helices of the protein, with little other secondary structure and no other tertiary structure. Relaxation kinetics studies were carried out to characterize the dynamics of core melting and formation in this protein. The unfolding and/or refolding response is induced by a laser-induced temperature jump between the folded and unfolded forms of E, and structural changes are monitored using the infrared amide I' absorbance at 1648-1651 cm(-1) that reports on the formation of solvent-protected, native-like helix in the core and by fluorescence emission changes from apomyoglobin's Trp14, a measure of burial of the indole group of this residue. The fluorescence kinetics data are monoexponential with a relaxation time of 14 micros. However, infrared kinetics data are best fit to a biexponential function with relaxation times of 14 and 59 micros. These relaxation times are very fast, close to the limits placed on folding reactions by diffusion. The 14 micros relaxation time is weakly temperature dependent and thus represents a pathway that is energetically downhill. The appearance of this relaxation time in both the fluorescence and infrared measurements indicates that this folding event proceeds by a concomitant formation of compact secondary and tertiary structures. The 59 micros relaxation time is much more strongly temperature dependent and has no fluorescence counterpart, indicating an activated process with a large energy barrier wherein nonspecific hydrophobic interactions between helix A and the G and H helices cause some helix burial but Trp14 remains solvent exposed. These results are best fit by a multiple-pathway kinetic model when U collapses to form the various folded core structures of E. Thus, the results suggest very robust dynamics for core formation involving multiple folding pathways and provide significant insight into the primary processes of protein folding.
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Hau, J., Andersson, E., & Carlsson, H. - E. (2001). Development and validation of a sensitive ELISA for quantification of secretory IgA in rat saliva and faeces. Laboratory Animals, 35(4), 301–306.
Abstract: Non-invasive measures of immunological markers are an attractive means of stress assessment in laboratory animals. Salivary IgA has been used successfully as a stress marker in the human, and several reports indicate the potential of secretory IgA as a non-invasive measure of stress in animals. The present paper describes the development of an ELISA using commercially available components for the quantification of rat IgA and validation of this assay for the quantification of rat secretory IgA in saliva and faeces. The concentration of IgA in rat saliva varied significantly between duplicate samples obtained from individual rats, and the viscosity and small total volume of rat saliva gave unsatisfactory results for IgA. Faecal IgA was present in high concentrations, and duplicate samples varied by only 2-3%. However, faecal IgA seemed less stable than IgA in other biological compartments, and this finding must be taken into consideration when using quantitative measurements of IgA as a marker of mucous humoral immune status.
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Ferguson, D. L., & Rosales-Ruiz, J. (2001). Loading the problem loader: the effects of target training and shaping on trailer-loading behavior of horses. J Appl Behav Anal, 34(4), 409–423.
Abstract: The purpose of this study was to develop an effective method for trailer loading horses based on principles of positive reinforcement. Target training and shaping were used to teach trailer-loading behavior to 5 quarter horse mares in a natural setting. All 5 had been trailer loaded before through the use of aversive stimulation. Successive approximations to loading and inappropriate behaviors were the dependent variables. After training a horse to approach a target, the target was moved to various locations inside the trailer. Horses started training on the left side of a two-horse trailer. After a horse was loading on the left side, she was moved to the right side, then to loading half on the right and half on the left. A limited-hold procedure and the presence of a companion horse seemed to facilitate training for 1 horse. Inappropriate behaviors fell to zero immediately after target training, and all the horses successfully completed the shaping sequence. Finally, these effects were observed to generalize to novel conditions (a different trainer and a different trailer).
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Virga, V., & Houpt, K. A. (2001). Prevalence of placentophagia in horses. Equine Vet J, 33(2), 208–210.
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Marlin, D. J., Schroter, R. C., White, S. L., Maykuth, P., Matthesen, G., Mills, P. C., et al. (2001). Recovery from transport and acclimatisation of competition horses in a hot humid environment. Equine Vet J, 33(4), 371–379.
Abstract: The aims of the present field-based study were to investigate changes in fit horses undergoing acclimatisation to a hot humid environment and to provide data on which to base recommendations for safe transport and acclimatisation. Six horses (age 7-12 years) were flown from Europe to Atlanta and underwent a 16 day period of acclimatisation. Exercise conditions during acclimatisation (wet bulb globe temperature index 27.6+/-0.0 [mean +/- s.e.]) were more thermally stressful compared with the European climate from which the horses had come (22.0+/-1.8, P<0.001). Following the flight, weight loss was 4.1+/-0.8% bodyweight and took around 7 days to recover. Water intake during the day was significantly increased (P<0.05) compared with night during acclimatisation. Daily mean exercise duration was 72+/-12 min and the majority of work was performed with a heart rate below 120 beats/min. Respiratory rate (fR) was increased (P<0.05) throughout acclimatisation compared with in Europe, but resting morning (AM) and evening (PM) rectal temperature (TREC), heart rate (fC) and plasma volume were unchanged. White blood cell (WBC) count was significantly increased at AM compared with in Europe on Days 4 and 10 of acclimatisation (P<0.01), but was not different by Day 16. In conclusion, horses exposed to hot humid environmental conditions without prior acclimatisation are able to accommodate these stresses and, with appropriate management, remain fit and clinically healthy, without significant risk of heat illness or heat-related disorders, provided they are allowed sufficient time to recover from transport, acclimatisation is undertaken gradually and they are monitored appropriately.
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Ricard, A., & Chanu, I. (2001). Genetic parameters of eventing horse competition in France. Genet Sel Evol, 33(2), 175–190.
Abstract: Genetic parameters of eventing horse competitions were estimated. About 13 000 horses, 30 000 annual results during 17 years and 110 000 starts in eventing competitions during 8 years were recorded. The measures of performance were logarithmic transformations of annual earnings, annual earnings per start, and annual earnings per place, and underlying variables responsible for ranks in each competition. Heritabilities were low (0.11 / 0.17 for annual results, 0.07 for ranks). Genetic correlations between criteria were high (greater than 0.90) except between ranks and earnings per place (0.58) or per start (0.67). Genetic correlations between ages (from 5 to 10 years old) were also high (more than 0.85) and allow selection on early performances. The genetic correlation between the results in different levels of competition (high/international and low/amateur) was near 1. Genetic correlations of eventing with other disciplines, which included partial aptitude needed for eventing, were very low for steeplechase races (0.18) and moderate with sport: jumping (0.45), dressage (0.58). The results suggest that selection on jumping performance will lead to some positive correlated response for eventing performance, but much more response could be obtained if a specific breeding objective and selection criteria were developed for eventing.
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Meershoek, L. S., Roepstorff, L., Schamhardt, H. C., Johnston, C., & Bobbert, M. F. (2001). Joint moments in the distal forelimbs of jumping horses during landing. Equine Vet J, 33(4), 410–415.
Abstract: Tendon injuries are an important problem in athletic horses and are probably caused by excessive loading of the tendons during demanding activities. As a first step towards understanding these injuries, the tendon loading was quantified during jump landings. Kinematics and ground reaction forces were collected from the leading and trailing forelimbs of 6 experienced jumping horses. Joint moments were calculated using inverse dynamic analysis. It was found that the variation of movement and loading patterns was small, both within and between horses. The peak flexor joint moments in the coffin and fetlock joints were larger in the trailing limb (-0.62 and -2.44 Nm/kg bwt, respectively) than in the leading limb (-0.44 and -1.93 Nm/kg bwt, respectively) and exceeded literature values for trot by 82 and 45%. Additionally, there was an extensor coffin joint moment in the first half of the stance phase of the leading limb (peak value 0.26+/-0.18 Nm/kg bwt). From these results, it was concluded that the loading of the flexor tendons during landing was higher in the trailing than in the leading limb and that there was an unexpected loading of the extensor tendon in the leading limb.
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Robert, C., Valette, J. P., & Denoix, J. M. (2001). The effects of treadmill inclination and speed on the activity of three trunk muscles in the trotting horse. Equine Vet J, 33(5), 466–472.
Abstract: The purpose of this study was to evaluate the effects of speed and slope on the activity of trunk muscles. The electromyographic (EMG) activity of the splenius (Sp), longissimus dorsi (LD) and rectus abdominis (RA) muscles was recorded with surface electrodes during treadmill locomotion at trot for different combinations of speed (3.5 to 6 m/s) and slope (0 to 6%). Raw EMG signals were processed to determine activity duration, onset and end and integrated EMG (IEMG). For the 3 muscles investigated, onset and end of activity were obtained earlier in the stride cycle when speed increased. A longer duration of activity for the LD, a shorter duration for the RA and an unchanged duration for the Sp were also observed. The IEMG of the latter was poorly affected by speed, whereas it increased linearly with speed for the 2 other muscles. When treadmill inclination changed from 0 to 6%, EMG activity of the LD and RA began and ended later; a longer activity duration was noted. Temporal parameters for Sp did not change with slope. A significant and progressive increase in the IEMG of the 3 muscles was observed with increasing slope. This evaluation of the activity of trunk muscles provides objective data for the use of speed or slope in training programmes.
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van der Kolk, J. H., Nachreiner, R. F., Schott, H. C., Refsal, K. R., & Zanella, A. J. (2001). Salivary and plasma concentration of cortisol in normal horses and horses with Cushing's disease. Equine Vet J, 33(2), 211–213.
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Malmgren, L., Andresen, Ø., & Dalin, A. - M. (2001). Effect of GnRH immunisation on hormonal levels, sexual behaviour, semen quality and testicular morphology in mature stallions. Equine vet. J., 33(1), 75–83.
Abstract: Summary The aim of this study was to investigate the effect of gonadotrophin-releasing hormone (GnRH) immunisation on mature stallions that had been used for breeding. Four Standardbred stallions were used in the study: 3 experimental animals and 1 control animal. Semen was collected regularly, i.e. twice/week, during the 4 months prior to the experimental period. The stallions were immunised against GnRH with a GnRH-BSA conjugate. Equimune was used as the adjuvant. The stallions were immunised on 5 occasions, 4 at 2 week intervals, and the fifth 4 weeks after the fourth. Blood samples were taken once a week for analysis of GnRH antibody titre and every third week for testosterone and oestrone sulphate analyses. Semen was collected once a week, and libido and sexual behaviour were observed. Ejaculate volume, sperm concentration, total number of sperm in the ejaculate, sperm motility and sperm morphology were evaluated. Testicular size was measured once a week. At the end of the study, the stallions were castrated, and a histological examination of the testes performed. All immunised stallions produced antibodies against GnRH, and plasma testosterone concentration decreased. However, the effect of immunisation varied between stallions. In 2 of the stallions, high levels of antibodies were found, while in the third, the level was moderate. Four weeks after the first immunisation, a decrease in libido was observed. Two months after the first immunisation, marked changes in semen quality were observed in the 2 stallions with high antibody titres. Fourteen weeks after the first immunisation, the total number of sperm/ejaculate had decreased from >8.6×109 to<2.7×109, sperm motility from >59 to<10% and the frequency of morphological normal spermatozoa had decreased from >60 to<14%. The dominating abnormalities were abnormal head shapes, proximal cytoplasmic droplets and detached heads. In the third stallion, only slight changes in semen quality were found. No changes were observed in the control stallion. Decreases in testicular size were noted in all of the experimental stallions. Pronounced histological alterations in the testes were observed in 2 of the stallions. It is concluded that the vaccine was effective in stimulating production of GnRH antibodies and in suppressing testicular function and androgen secretion. However, there was an individual variation in the responses among the stallions and, further, libido was not totally suppressed.
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