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Author |
Polverini, E.; Cugini, G.; Annoni, F.; Abbruzzetti, S.; Viappiani, C.; Gensch, T. |
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Title |
Molten globule formation in apomyoglobin monitored by the fluorescent probe Nile Red |
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Journal Article |
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Year |
2006 |
Publication |
Biochemistry |
Abbreviated Journal |
Biochemistry |
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Volume |
45 |
Issue |
16 |
Pages |
5111-5121 |
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Keywords |
Animals; Apoproteins/*chemistry/*metabolism; Binding Sites; Computer Simulation; Fluorescent Dyes/analysis; Horses; Hydrogen-Ion Concentration; Models, Molecular; Myoglobin/*chemistry/*metabolism; Oxazines/*analysis/chemistry; Protein Binding; Protein Folding; Protein Structure, Tertiary |
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Abstract |
The interaction of nile red (NR) with apomyoglobin (ApoMb) in the native (pH 7) and molten globule (pH 4) states was investigated using experimental and computational methods. NR binds to hydrophobic locations in ApoMb with higher affinity (K(d) = 25 +/- 5 microM) in the native state than in the molten globule state (K(d) = 52 +/- 5 microM). In the molten globule state, NR is located in a more hydrophobic environment. The dye does not bind to the holoprotein, suggesting that the binding site is located at the heme pocket. In addition to monitoring steady-state properties, the fluorescence emission of NR is capable of tracking submillisecond, time-resolved structural rearrangements of the protein, induced by a nanosecond pH jump. Molecular dynamics simulations were run on ApoMb at neutral pH and at pH 4. The structure obtained for the molten globule state is consistent with the experimentally available structural data. The docking of NR with the crystal structure shows that the ligand binds into the binding pocket of the heme group, with an orientation bringing the planar ring system of NR to overlap with the position of two of the heme porphyrin rings in Mb. The docking of NR with the ApoMb structure at pH 4 shows that the dye binds to the heme pocket with a slightly less favorable binding energy, in keeping with the experimental K(d) value. Under these conditions, NR is positioned in a different orientation, reaching a more hydrophobic environment in agreement with the spectroscopic data. |
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Dipartimento di Fisica, Universita degli Studi di Parma, Viale G. P. Usberti 7/A, 43100 Parma, Italy |
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0006-2960 |
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PMID:16618100 |
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Equine Behaviour @ team @ |
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3763 |
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Author |
Balakrishnan, G.; Hu, Y.; Spiro, T.G. |
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Title |
Temperature-jump apparatus with Raman detection based on a solid-state tunable (1.80-2.05 microm) kHz optical parametric oscillator laser |
Type |
Journal Article |
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Year |
2006 |
Publication |
Applied Spectroscopy |
Abbreviated Journal |
Appl Spectrosc |
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Volume |
60 |
Issue |
4 |
Pages |
347-351 |
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Keywords |
Animals; Cytochromes c/analysis; Horses; Lasers; Myoglobin/metabolism; Spectrum Analysis, Raman/*instrumentation/*methods; *Temperature |
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The operating characteristics of a pulsed (10 ns) tunable near-infrared (NIR) laser source are described for temperature-jump (T-jump) applications. A Q-switched Nd:YLF laser (approximately 10 ns pulses) with a 1 kHz repetition rate is used to pump a potassium titanyl arsenate (KTA) crystal-based optical parametric oscillator (OPO), producing approximately 1 mJ NIR pulses that are tunable (1.80-2.05 microm) across the 1.9 microm vibrational overtone band of water. This T-jump source has been coupled to a deep ultraviolet (UV) probe laser for Raman studies of protein dynamics. T-jumps of up to 30 degrees C, as measured via the O-H stretching Raman band of water, are readily achieved. Application to cytochrome c unfolding is demonstrated. |
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Department of Chemistry, Princeton University, Princeton, New Jersey 08544, USA |
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0003-7028 |
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PMID:16613628 |
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Equine Behaviour @ team @ |
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3764 |
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Murray, J.K.; Singer, E.R.; Morgan, K.L.; Proudman, C.J.; French, N.P. |
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Title |
The risk of a horse-and-rider partnership falling on the cross-country phase of eventing competitions |
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Journal Article |
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Year |
2006 |
Publication |
Equine Veterinary Journal |
Abbreviated Journal |
Equine Vet J |
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Volume |
38 |
Issue |
2 |
Pages |
158-163 |
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Keywords |
*Accidental Falls; Animals; Athletic Injuries/epidemiology/*veterinary; Case-Control Studies; Horses/*injuries; Humans; Logistic Models; Physical Conditioning, Animal; Prospective Studies; Risk Factors; Safety; Sports; Time Factors |
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Abstract |
REASONS FOR PERFORMING STUDY: Fatalities resulting from horse falls occurring during the cross-country phase of eventing competitions initiated epidemiological investigation of the risk factors associated with horse falls. OBJECTIVES: To identify variables that increased or decreased the risk of a horse fall during the cross-country phase of an eventing competition. METHODS: Data were collected from randomly selected British Eventing competitions held in Great Britain during 2001 and 2002. Data were obtained for 173 cases (jumping efforts resulting in a fall of the horse-and-rider partnership) and 503 matched controls (jumping efforts not resulting in a fall). The risk of falling was modelled using conditional logistic regression. RESULTS: An increased risk of a horse fall was associated with jumping into or out of water; taking off from good-to-soft, soft or heavy ground; fences with a drop landing; nonangled fences with a spread > or =2 m; and angled fences. Other risk factors included riders who knew that they were in the lead within the competition before the cross-country phase; an inappropriate speed of approach to the fence (too fast or too slow); horse-and-rider partnerships that had not incurred refusals at earlier fences; and riders who received cross-country tuition. CONCLUSIONS: This study has identified modifiable course- and fence-level risk factors for horse falls during the cross-country phase of eventing competitions. The risk of horse and rider injury at eventing competitions should be reduced by 3 simple measures; maintaining good to firm take-off surfaces at fences, reducing the base spread of fences to <2 m and reducing the use of fences at which horses are required to jump into or out of water. Risk reduction arising from course and fence modification needs to be confirmed by intervention studies. Potential relevance: Knowledge of factors that increase or decrease the risk of a horse fall can be used by UK governing bodies of the sport to reduce the risk of horse falls on the cross-country phase of eventing competitions, and reduce the risk of horse and rider injuries and fatalities. As one in 3 horses that fall injure themselves and one in 100 horse falls results in fatality to the horse, we suggest that immediate consideration is given to these recommendations. |
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Epidemiology Group, Faculty of Veterinary Science, University of Liverpool, Leahurst, Chester High Road, Neston, South Wirral CH64 7TE, UK |
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0425-1644 |
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PMID:16536386 |
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Call Number |
Equine Behaviour @ team @ |
Serial |
3765 |
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Author |
Hurn, S.D.; Turner, A.G. |
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Title |
Ophthalmic examination findings of Thoroughbred racehorses in Australia |
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Journal Article |
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Year |
2006 |
Publication |
Veterinary Ophthalmology |
Abbreviated Journal |
Vet Ophthalmol |
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Volume |
9 |
Issue |
2 |
Pages |
95-100 |
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Keywords |
Animals; Cataract/epidemiology/veterinary; Diagnostic Techniques, Ophthalmological/veterinary; Eye Diseases/diagnosis/epidemiology/*veterinary; Female; Horse Diseases/diagnosis/*epidemiology; Horses; Male; Prevalence; Victoria/epidemiology |
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Abstract |
OBJECTIVE: To record the prevalence and document the types of eye disease in population of Thoroughbred racehorses in Victoria, Australia. DESIGN: Prospective study. ANIMALS: Two hundred four Thoroughbred racehorses. PROCEDURE: All horses and both eyes were examined at four metropolitan and two country racing stable complexes. Ophthalmic exam was performed following dark adaptation with a transilluminator, biomicroscope, and direct ophthalmoscope. Intraocular pressures were measured when indicated. Both pupils were dilated with tropicamide when indicated. RESULTS: One hundred eighty-two (89.2%) flat-racing and 22 (10.8%) jump-racing (hurdle or steeple) horses were examined. Age range: 2-9 years (mean 3.7 years, median 3); 97 (47.5%) male-neuter, 74 (36. 3%) female, 33 (16.2%) male. Potential vision-threatening eye disease was present in 15 (7.4%) different horses: complete lenticular cataracts 3, posterior lens luxation and cataract 1, large peripapillary 'butterfly' inactive lesions 3, large peripapillary 'butterfly' active lesions 2, peripapillary focal inactive 'bullet hole' chorioretinal lesions (> 20) 5, optic nerve atrophy 1. Non-vision threatening eye disease was present in 117 (57.4%) different horses, involving one or more ocular structures: lower eyelid scars 3; periocular fibropapillomatous disease 1; third eyelid squamous cell carcinoma 1; corneal scars 6; corneal band opacity 2; anterior iris synechia 1; developmental cataracts 36 (17.2%); peripapillary focal inactive 'bullet hole' chorioretinal lesions (< 20) 103 (50.0%); linear peripapillary hyperpigmentation bands 16 (7.9%). Unusual variations of normal ocular anatomy and colobomata was recorded in 11 (5.4%) different horses: granular iridica hypoplasia 3, granular iridica hyperplasia 2, multilobular granular iridica cyst 1, microcornea 1, hyaloid remnant 1, rotated optic nerve head 1, coloboma of the lens 1, atypical coloboma of the retina 1. CONCLUSIONS: This survey demonstrates that the prevalence of vision-threatening eye disease in racing horses may be greater than previously perceived, and highlights the importance of ocular examination within any routine physical examination of horses. |
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All Animal Eye Services, Mount Waverley, Victoria, Australia. sdhurn@bigpond.com |
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1463-5216 |
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PMID:16497233 |
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Call Number |
Equine Behaviour @ team @ |
Serial |
3766 |
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Author |
Bykov, S.; Lednev, I.; Ianoul, A.; Mikhonin, A.; Munro, C.; Asher, S.A. |
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Title |
Steady-state and transient ultraviolet resonance Raman spectrometer for the 193-270 nm spectral region |
Type |
Journal Article |
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Year |
2005 |
Publication |
Applied Spectroscopy |
Abbreviated Journal |
Appl Spectrosc |
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Volume |
59 |
Issue |
12 |
Pages |
1541-1552 |
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Keywords |
Animals; Equipment Design; Equipment Failure Analysis; Horses; Kinetics; Metmyoglobin/*analysis; Myocardium/*metabolism; Reproducibility of Results; Sensitivity and Specificity; Spectrophotometry, Ultraviolet/*instrumentation/methods; Spectrum Analysis, Raman/*instrumentation/methods |
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Abstract |
We describe a state-of-the-art tunable ultraviolet (UV) Raman spectrometer for the 193-270 nm spectral region. This instrument allows for steady-state and transient UV Raman measurements. We utilize a 5 kHz Ti-sapphire continuously tunable laser (approximately 20 ns pulse width) between 193 nm and 240 nm for steady-state measurements. For transient Raman measurements we utilize one Coherent Infinity YAG laser to generate nanosecond infrared (IR) pump laser pulses to generate a temperature jump (T-jump) and a second Coherent Infinity YAG laser that is frequency tripled and Raman shifted into the deep UV (204 nm) for transient UV Raman excitation. Numerous other UV excitation frequencies can be utilized for selective excitation of chromophoric groups for transient Raman measurements. We constructed a subtractive dispersion double monochromator to minimize stray light. We utilize a new charge-coupled device (CCD) camera that responds efficiently to UV light, as opposed to the previous CCD and photodiode detectors, which required intensifiers for detecting UV light. For the T-jump measurements we use a second camera to simultaneously acquire the Raman spectra of the water stretching bands (2500-4000 cm(-1)) whose band-shape and frequency report the sample temperature. |
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Department of Chemistry, University of Pittsburgh, Pittsburgh, Pennsylvania 15260, USA |
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0003-7028 |
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PMID:16390595 |
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Equine Behaviour @ team @ |
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3767 |
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Author |
Young, L.E.; Rogers, K.; Wood, J.L.N. |
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Title |
Left ventricular size and systolic function in Thoroughbred racehorses and their relationships to race performance |
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Journal Article |
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Year |
2005 |
Publication |
Journal of Applied Physiology (Bethesda, Md. : 1985) |
Abbreviated Journal |
J Appl Physiol |
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Volume |
99 |
Issue |
4 |
Pages |
1278-1285 |
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Keywords |
*Adaptation, Physiological; Aging/physiology; Animal Husbandry; Animals; *Echocardiography; Female; Heart/*physiology; Heart Ventricles; Horses/*physiology; Male; *Physical Conditioning, Animal; Running/*physiology; Stroke Volume; Systole; Task Performance and Analysis |
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Cardiac morphology in human athletes is known to differ, depending on the sports-specific endurance component of their events, whereas anecdotes abound about superlative athletes with large hearts. As the heart determines stroke volume and maximum O(2) uptake in mammals, we undertook a study to test the hypothesis that the morphology of the equine heart would differ between trained horses, depending on race type, and that left ventricular size would be greatest in elite performers. Echocardiography was performed in 482 race-fit Thoroughbreds engaged in either flat (1,000-2,500 m) or jump racing (3,200-6,400 m). Body weight and sex-adjusted measures of left ventricular size were largest in horses engaged in jump racing over fixed fences, compared with horses running shorter distances on the flat (range 8-16%). The observed differences in cardiac morphologies suggest that subtle differences in training and competition result in cardiac adaptations that are appropriate to the endurance component of the horses' event. Derived left ventricular mass was strongly associated with published rating (quality) in horses racing over longer distances in jump races (P < or = 0.001), but less so for horses in flat races. Rather, left ventricular ejection fraction and left ventricular mass combined were positively associated with race rating in older flat racehorses running over sprint (<1,408 m) and longer distances (>1,408 m), explaining 25-35% of overall variation in performance, as well as being closely associated with performance in longer races over jumps (23%). These data provide the first direct evidence that cardiac size influences athletic performance in a group of mammalian running athletes. |
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Centre for Equine Studies, Animal Health Trust, Newmarket, Suffolk, UK. lesley.young@aht.org.uk |
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8750-7587 |
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PMID:15920096 |
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Equine Behaviour @ team @ |
Serial |
3768 |
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Author |
Rollot, Y.; Lecuyer, E.; Chateau, H.; Crevier-Denoix, N. |
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Title |
Development of a 3D model of the equine distal forelimb and of a GRF shoe for noninvasive determination of in vivo tendon and ligament loads and strains |
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Journal Article |
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Year |
2004 |
Publication |
Equine Veterinary Journal |
Abbreviated Journal |
Equine Vet J |
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Volume |
36 |
Issue |
8 |
Pages |
677-682 |
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Keywords |
Animals; Biomechanics; Floors and Floorcoverings; Forelimb/*physiology/ultrasonography; Gait/physiology; Horses/*physiology; Image Processing, Computer-Assisted; Imaging, Three-Dimensional/methods/*veterinary; Ligaments, Articular/*physiology; Locomotion/*physiology; Models, Biological; Shoes; Tendons/*physiology; Toe Joint/physiology/ultrasonography |
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REASONS FOR PERFORMING STUDY: As critical locomotion events (e.g. high-speed and impacts during racing, jump landing) may contribute to tendinopathies, in vivo recording of gaits kinematic and dynamic parameters is essential for 3D reconstruction and analysis. OBJECTIVE: To propose a 3D model of the forelimb and a ground reaction force recording shoe (GRF-S) for noninvasively quantifying tendon and ligament loads and strains. METHODS: Bony segments trajectories of forelimbs placed under a power press were recorded using triads of ultrasonic kinematic markers linked to the bones. Compression cycles (from 500-6000 N) were applied for different hoof orientations. Locations of tendon and ligament insertions were recorded with regard to the triads. The GRF-S recorded GRF over the hoof wall and used four 3-axis force sensors sandwiched between a support shoe and the shoe to be tested. RESULTS: Validation of the model by comparing calculated and measured superficial digital flexor tendon strains, and evaluation of the role of proximal interphalangeal joint in straight sesamoidean ligament and oblique sesamoidean ligament strains, were successfully achieved. Objective comparisons of the 3 components of GRF over the hoof for soft and hard grounds could be recorded, where the s.d. of GRF norm was more important on hard ground at walk and trot. CONCLUSIONS: Soft grounds (sand and rubber) dissipate energy by lowering GRF amplitude and diminish bounces and vibrations at impact. At comparable speed, stance phase was longer on soft sand ground. POTENTIAL RELEVANCE: The conjugate use of the GRF-S and the numerical model would help to quantify and analyse ground/shoe combination on comfort, propulsion efficiency or lameness recovery. |
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UMR INRA-ENVA de Biomecanique et Pathologie Locomotrice du Cheval, Ecole Nationale Veterinaire d'Alfort, 7, Avenue du General de Gaulle, 94704 Maisons-Alfort, France |
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0425-1644 |
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PMID:15656495 |
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Equine Behaviour @ team @ |
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3769 |
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Author |
Abbruzzetti, S.; Viappiani, C.; Sinibaldi, F.; Santucci, R. |
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Title |
Kinetics of histidine dissociation from the heme Fe(III) in N-fragment (residues 1-56) of cytochrome c |
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Journal Article |
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Year |
2004 |
Publication |
The Protein Journal |
Abbreviated Journal |
Protein J |
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Volume |
23 |
Issue |
8 |
Pages |
519-527 |
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Keywords |
Animals; Cytochromes c/*chemistry; Enzyme Activation; Histidine/*chemistry; Horses; Hydrogen-Ion Concentration; Kinetics; Lasers; Ligands; Peptide Mapping; Photolysis; Spectrophotometry |
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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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Dipartimento di Fisica, Universita degli Studi di Parma, Parco Area delle Scienze 7/A 43100 Parma, Italy |
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1572-3887 |
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PMID:15648974 |
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Call Number |
Equine Behaviour @ team @ |
Serial |
3770 |
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Author |
Santamaria, S.; Bobbert, M.E.; Back, W.; Barneveld, A.; van Weeren, P.R. |
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Title |
Variation in free jumping technique within and among horses with little experience in show jumping |
Type |
Journal Article |
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Year |
2004 |
Publication |
American Journal of Veterinary Research |
Abbreviated Journal |
Am J Vet Res |
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Volume |
65 |
Issue |
7 |
Pages |
938-944 |
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Keywords |
*Acceleration; Analysis of Variance; Animals; Biomechanics; Forelimb/physiology; Gait/*physiology; Hindlimb/physiology; Horses/*physiology; Locomotion/*physiology; Models, Biological; Video Recording |
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Abstract |
OBJECTIVE: To quantify variation in the jumping technique within and among young horses with little jumping experience, establish relationships between kinetic and kinematic variables, and identify a limited set of variables characteristic for detecting differences in jumping performance among horses. ANIMALS: Fifteen 4-year-old Dutch Warmblood horses. PROCEDURE: The horses were raised under standardized conditions and trained in accordance with a fixed protocol for a short period. Subsequently, horses were analyzed kinematically during free jumping over a fence with a height of 1.05 m. RESULTS: Within-horse variation in all variables that quantified jumping technique was smaller than variation among horses. However, some horses had less variation than others. Height of the center of gravity (CG) at the apex of the jump ranged from 1.80 to 2.01 m among horses; this variation could be explained by the variation in vertical velocity of the CG at takeoff (r, 0.78). Horses that had higher vertical velocity at takeoff left the ground and landed again farther from the fence, had shorter push-off phases for the forelimbs and hind limbs, and generated greater vertical acceleration of the CG primarily during the hind limb push-off. However, all horses cleared the fence successfully, independent of jumping technique. CONCLUSIONS AND CLINICAL RELEVANCE: Each horse had its own jumping technique. Differences among techniques were characterized by variations in the vertical velocity of the CG at takeoff. It must be determined whether jumping performance later in life can be predicted from observing free jumps of young horses. |
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Address |
Department of Equine Sciences, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 12, NL-3584 CM Utrecht, The Netherlands |
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0002-9645 |
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PMID:15281652 |
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Equine Behaviour @ team @ |
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3772 |
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Author |
Pinchbeck, G.L.; Clegg, P.D.; Proudman, C.J.; Morgan, K.L.; French, N.P. |
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Case-control investigation of the factors affecting the risk of horses falling during steeplechase racing in the UK |
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Journal Article |
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2004 |
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The Veterinary Record |
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Vet. Rec. |
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155 |
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1 |
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11-15 |
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Accidental Falls/*prevention & control/*statistics & numerical data; Animals; Athletic Injuries/epidemiology/etiology/prevention & control/*veterinary; Case-Control Studies; England/epidemiology; Horses/*injuries; Risk Factors; Running/*injuries |
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Abstract |
A concurrent case-control study of 12 UK racecourses was made between March 1, 2000, and August 31, 2001, to identify and quantify the factors associated with the risk of horses falling in steeplechase races. Cases were defined as a jumping effort at a steeplechase fence that resulted in a fall and controls were defined as a successful jumping effort over any steeplechase fence at any of the 12 racecourses within 14 days before or after the case fall. Information on the horse, the jockey and the race were collected and all the fences on all the courses were surveyed. Conditional logistic regression was used to examine the relationships between the predictor variables and the risk of falling. There was one fall per 254 jumping efforts. The risk of a horse falling decreased the more times it had raced on a particular racecourse. The number of fences, the distance from the previous fence and the nature of the previous fence also affected the risk of falling. If the previous fence was a water jump the risk of falling increased; fences that were sited on flat or slight uphill gradients (up to approximately 1 in 25) were associated with a lower risk of horses falling than downhill fences, and higher takeoff boards were associated with a higher risk of falling. |
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Department of Veterinary Clinical Science, University of Liverpool, Leahurst, Neston CH64 7TE |
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0042-4900 |
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PMID:15264483 |
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Call Number |
Equine Behaviour @ team @ |
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3773 |
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