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Author |
Steinhoff, H.J. |
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Title |
A continuous wave laser T-jump apparatus and its application to chemical reactions in hemoglobin single crystals |
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Journal Article |
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Year |
1988 |
Publication |
Journal of Biochemical and Biophysical Methods |
Abbreviated Journal |
J Biochem Biophys Methods |
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Volume |
15 |
Issue |
6 |
Pages |
319-330 |
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Keywords |
Animals; Chemistry; Crystallization; *Heat; *Hemoglobins; Horses/blood; *Lasers; Methemoglobin; Solutions; Thermodynamics; Thiocyanates |
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Abstract |
A laser temperature jump apparatus is constructed where the T-jump is achieved by means of the direct absorption of continuous laser radiation of low intensity by a solid sample. The final temperature in the irradiated volume element is reached when the absorbed radiation power equals the dissipation of heat by heat conduction. The time range from the beginning of irradiation to the stationary state depends on the geometry of the irradiated volume element and is less than 10 ms. The heating laser beam is simultaneously used to detect the relaxation to the new chemical equilibrium in the sample. Relaxation processes with relaxation rates between 10(2) s-1 and less than 10(-3) s-1 on samples with volumes less than 10(-3) mm3 may be investigated using this T-jump method. One application of this method is the determination of reaction rates of ligand reactions in hemoglobin single crystals. Rate constants obtained for the reaction of thiocyanate with crystallized horse methemoglobin are presented. |
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Institut fur Biophysik, Ruhr-Universitat Bochum, F.R.G |
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0165-022X |
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PMID:3379245 |
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Equine Behaviour @ team @ |
Serial |
3804 |
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Author |
Dyson, H.J.; Beattie, J.K. |
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Title |
Spin state and unfolding equilibria of ferricytochrome c in acidic solutions |
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Journal Article |
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Year |
1982 |
Publication |
The Journal of Biological Chemistry |
Abbreviated Journal |
J Biol Chem |
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Volume |
257 |
Issue |
5 |
Pages |
2267-2273 |
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Keywords |
Animals; *Cytochrome c Group; Electron Spin Resonance Spectroscopy; Heme; Horses; Hydrogen-Ion Concentration; Kinetics; Ligands; Myocardium; Protein Binding; Protein Conformation; Spectrophotometry; Temperature |
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Abstract |
Equilibrium, stopped flow, and temperature-jump spectrophotometry have been used to identify processes in the unfolding of ferricytochrome c in acidic aqueous solutions. A relaxation occurring in approximately 100 microseconds involves perturbation of a spin-equilibrium between two folded conformers of the protein with methionine-80 coordinated or dissociated from the heme iron. The protein unfolds more slowly, in milliseconds, with dissociation and protonation of histidine-18. These two transitions appear cooperative in equilibrium measurements at low (0.01 M) ionic strength, but are separated at higher (0.10 M) ionic strength. They are resolved under both conditions in the dynamic measurements. The spin-equilibrium description permits a unified explanation of a number of properties of ferricytochrome c in acidic aqueous solutions. |
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0021-9258 |
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PMID:6277891 |
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Equine Behaviour @ team @ |
Serial |
3807 |
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Author |
Czerlinski, G.H.; Erickson, J.O.; Theorell, H. |
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Title |
Chemical relaxation studies on the horse liver alcohol dehydrogenase system |
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Journal Article |
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Year |
1979 |
Publication |
Physiological Chemistry and Physics |
Abbreviated Journal |
Physiol Chem Phys |
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Volume |
11 |
Issue |
6 |
Pages |
537-569 |
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Keywords |
Alcohol Oxidoreductases/*metabolism; Animals; Buffers; Electron Transport; Ethanol/metabolism; Horses; Hydrogen-Ion Concentration; Liver/*enzymology; Mathematics; NAD/metabolism; Oscillometry; Osmolar Concentration; Temperature; Time Factors |
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Chemical relaxation studies on the system horse liver alcohol dehydrogenase, nicotinamide adenine dinucleotide, and ethanol were conducted observing fluorescence changes between 400 and 500 nm. Temperature-jump experiments were performed at pH 6.5, 7.0, 8.0, and 9.0; concentration-jump experiments at pH 9.0. The reciprocal of the slowest relaxation time was found to be linearly dependent upon the enzyme concentration for relatively low enzyme concentrations, as predicted earlier. Use of the wide pH-range necessitated expression of the four apparent dissociation constants of the catalytic reaction cycle in terms of pH-independent constants. The system was described in terms of only one (or two) catalysis-linked protons not associated with the electron transfer. Protonic steps in a buffered system are in rapid equilibrium, too fast to be measured with the equipment available. Assuming only two of the four bimolecular reaction steps in the four-step cycle are fast compared to the remaining two, six cases may be considered with six expressions for the reciprocal of the slowest relaxation time. Comparison with the experimental data revealed that the bimolecular reaction steps governing the slowest relaxation time change with pH. Above the effective time resolution of the temperature-lump instrument with fluorescence detection (0.1 msec) only one other relaxation time was detectable and only at pH 9. This relaxation time, found to be independent of the concentration of all reactants within experimental error (r = 10 +/- 5 msec), is most likely due to an interconversion among ternary complexes. |
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0031-9325 |
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PMID:44918 |
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Equine Behaviour @ team @ |
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3813 |
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Author |
Hasumi, H. |
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Title |
Kinetic studies on isomerization of ferricytochrome c in alkaline and acid pH ranges by the circular dichroism stopped-flow method |
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Journal Article |
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Year |
1980 |
Publication |
Biochimica et Biophysica Acta |
Abbreviated Journal |
Biochim Biophys Acta |
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Volume |
626 |
Issue |
2 |
Pages |
265-276 |
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Circular Dichroism; *Cytochrome c Group; Hydrogen-Ion Concentration; Isomerism; Kinetics; Spectrophotometry |
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The isomerization of horse-heart ferricytochrome c caused by varying pH was kinetically studied by using circular dichroism (CD) and optical absorption stopped-flow techniques. In the pH range of 7--13, the existence of the three different forms of ferricytochrome c (pH less than 10, pH 10--12, and pH greater than 12) was indicated from the statistical difference CD spectra. On the basis of analyses of the stopped-flow traces in the near-ultraviolet and Soret wavelength regions, the isomerization of ferricytochrome c from neutral form to the above three alkaline forms was interpreted as follows (1) below pH 10, the replacement of the intrinsic ligand of methionine residue by lysine residue occurs; (2) between pH 10 and 12, the uncoupling of the polypeptide chain from close proximity of the heme group occurs first, followed by the interconversion of the intrinsic ligands; and (3) above pH 12, hydroxide form of ferricytochrome c is formed, though the replacement of the intrinsic ligand by extrinsic ligands may occur via different routes from those below pH 12. The CD changes at 288 nm and in the Soret region caused by the pH-jump (down) from pH 6.0 to 1.6 were compared with the appearance of the 620-nm absorption band ascribed to the formation of the high-spin form of ferricytochrome c. Both CD and absorption changes indicated that the isomerization at pH 1.6 consisted of two processes: one proceeded within the dead-time (about 2 ms) of the stopped-flow apparatus and the other proceeded at a determinable rate with the apparatus. On the basis of these results, the isomerization of ferricytochrome c at pH 1.6 was explained as follows: (1) the transition from the low-spin form to the high-spin forms occurs within about 2 ms, the dead-time of the stopped-flow apparatus; and (2) the polypeptide chain is unfolded after the formation of the high-spin form. |
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0006-3002 |
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Notes |
PMID:6260152 |
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Call Number |
refbase @ user @ |
Serial |
3876 |
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Author |
Czerlinski, G.H.; Wagner, M.; Erickson, J.O.; Theorell, H. |
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Title |
Chemical relaxation studies on the system liver alcohol dehydrogenase, NADH and imidazole |
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Journal Article |
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Year |
1975 |
Publication |
Acta Chemica Scandinavica. Series B: Organic Chemistry and Biochemistry |
Abbreviated Journal |
Acta Chem Scand B |
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29 |
Issue |
8 |
Pages |
797-810 |
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Alcohol Oxidoreductases/*metabolism; Animals; Computers; Hydrogen-Ion Concentration; Imidazoles/*metabolism; Kinetics; Liver/enzymology/*metabolism; Mathematics; Models, Chemical; NAD/*metabolism; Time Factors |
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Several years ago, Theorell and Czerlinski conducted experiments on the system of horse liver alcohol dehydrogenase, reduced nicotinamide adenine dinucleotide and imidazole, using the first version of the temperature jump apparatus with detection of changes in fluorescence. These early experiments were repeated with improved instrumentation and confirmed the early experiments in general terms. However, the improved detection system allowed to measure a slight concentration dependence of the relaxation time of around 3 ms. Furthermore, the chemical relaxation time was smaller than the one determined earlier (by factor 2). The data were evaluated much more rigorously than before, allowing an appropriate interpretation of the results. The observed relaxation time is largely due to rate constants in an interconversion of ternary complexes, which are faster than three (of the four) dissociation rate constants, determined previously by Theorell and McKinley-McKee.1,2 This fact contributed to earlier difficulties of finding any concentration dependence. However, the binding of imidazole to the binary enzyme-coenzyme complex can be made to couple kinetically into the interconversion rate of the two ternary complexes. The observed signal derives largely from the ternary complex(es). A substantial fluorescence signal change is associated with the observed relaxation process, suggesting a relocation of the imidazole in reference to the nicotinamide moiety of the bound coenzyme. Nine models are considered with two types of coupling of pre-equilibria (none-all). Quantitative evaluations favor the model with two ternary complexes connected by an interconversion outside the four-step (bimolecular) cycle. The ternary complex outside the cycle has much higher fluorescence yield than the one inside. The interconversion equilibrium is near unity for imidazole. If it would be shifted very much to the side of the “dead-end” complex (as in isobutyramide?!), stimulating action could not take place. |
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0302-4369 |
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PMID:882 |
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refbase @ user @ |
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3887 |
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Ducro, B.J.; Koenen, E.P.C.; van Tartwijk, J.M.F.M.; Bovenhuis, H. |
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Title |
Genetic relations of movement and free-jumping traits with dressage and show-jumping performance in competition of Dutch Warmblood horses |
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Journal Article |
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2007 |
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Livestock Science |
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107 |
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2-3 |
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227-234 |
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Horse; Heritability; Movement; Free-jumping; Dressage |
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Genetic parameters for traits evaluated at the studbook entry inspection and genetic correlations with dressage and show-jumping performance in competition were estimated. Data comprised 36,649 Warmblood horses that entered the studbook between 1992 and 2002. The genetic analyses were performed using univariate and bivariate animal models. Heritabilities of the studbook entry traits were estimated in the range 0.15-0.40. The movement traits showed moderate to strong mutual genetic correlations, whereas the genetic correlations of movement traits with free-jumping traits were weak to moderate. The free-jumping traits showed strong to very strong mutual genetic correlations. Competition results of 33,459 horses with performance in dressage and 30,474 horse with performance in show-jumping were linked to the studbook entry data to estimate the genetic relationship with performance in competition. Heritability estimates for dressage and show jumping were 0.14. Genetic correlations of the movement traits with dressage were moderate to strong, and with show-jumping weak to moderate. Genetic correlations of the free-jumping traits with dressage were weak to moderate and unfavourable. The free-jumping traits were genetically strong to very strong correlated to show-jumping. It was concluded that a selection of the traits evaluated at the studbook entry inspection will favourably contribute to estimation of breeding values for sport performance. |
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refbase @ user @ |
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3947 |
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Buchner, H.H.F.; Obermuller, S.; Scheidl, M. |
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Title |
Body Centre of Mass Movement in the Sound Horse |
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Journal Article |
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2000 |
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The Veterinary Journal |
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160 |
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3 |
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225-234 |
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Horse; centre of mass; kinematics; segment model; locomotion. |
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The body centre of mass (BCM) is a key factor in the analysis of equine locomotion, as its position and movement determines the distribution and magnitude of loads on the limbs. In this study, the three-dimensional (3D) movement of the BCM in walking and trotting horses was assessed using a kinematic, segmental method. Thirty markers representing 20 body segments were recorded in 12 sound horses while standing, walking and trotting on a treadmill using a high-speed video system. Based on segmental inertial data, 3D positions of the segmental centres of mass as well as the total BCM were calculated. The position within the trunk during square standing and the movements of the BCM were determined for the three planes. The position of the BCM in the standing horse is presented relative to external reference points. At the trot, vertical displacement amplitude of the BCM amounted to 53 (6) mm as mean (sd), which was 27% smaller than external trunk movement. Medio-lateral displacement amplitude of the BCM was 19 (4) mm, 34% less than trunk amplitude. Sagittal forward-backward oscillations of the BCM independent from general forward movement were 13 (3) mm, being 24% less than trunk movements. At the walk, vertical, medio-lateral and sagittal BCM movements were smaller than trunk movements by 43, 65 and 65% respectively. The results show reduced and efficient BCM movements compared to the trunk and form a basis for the assessment of various clinical conditions such as lameness, the influence of a rider and various dressage performances. |
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refbase @ user @ |
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3959 |
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Hodson, E.F.; Clayton, H.M.; Lanovaz, J.L. |
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Temporal analysis of walk movements in the Grand Prix dressage test at the 1996 Olympic Games |
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Journal Article |
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1999 |
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Applied Animal Behaviour Science |
Abbreviated Journal |
Appl. Anim. Behav. Sci. |
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62 |
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2-3 |
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89-97 |
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Dressage; Horse; Kinematics; Locomotion; Gait |
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Video analysis was used to measure temporal characteristics of the collected walk, extended walk and half pirouette at walk of eleven competitors during the team dressage competition at the 1996 Summer Olympic Games in Atlanta, GA. Forelimb stance durations, hind limb stance durations, lateral step intervals and diagonal step intervals were symmetrical for the right and left sides in the collected and extended walk strides, but there were left-right asymmetries in the forelimb stance duration and in the lateral step interval in the half pirouette strides. For both collected and extended walk strides, hind limb stance duration was significantly longer than forelimb stance duration. The mean values for the group of eleven horses showed that the collected and extended walks had a regular rhythm. The half pirouette strides showed an irregularity in which there was a short interval between footfalls of the outside forelimb and inside hind limb, and along interval between footfalls of the inside hind limb and inside forelimb. This irregularity reflected an early placement of the inside hind limb. The stance times of both hind limbs were prolonged and this finding, in combination with the early placement of the inside hind limb, led to an increase in the period of tripedal support in each stride of the half pirouette. This was interpreted as a means of maintaining the horses' balance in the absence of forward movement. |
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refbase @ user @ |
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3960 |
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Huizinga, H.A.; van der Werf, J.H.J.; Korver, S.; van der Meij, G.J.W. |
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Stationary performance testing of stallions from the Dutch Warmblood riding horse population. 1. Estimated genetic parameters of scored traits and the genetic relation with dressage and jumping competition from offspring of breeding stallions |
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1991 |
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Livestock Production Science |
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27 |
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2-3 |
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231-244 |
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dressage; genetic parameters; horse; jumping; performance; stallion |
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The stationary performance testing (SPT) of stallions as breeding candidates in the Dutch Warmblood riding horse population is evaluated. Genetic and phenotypic parameters of traits scored during SPT and the genetic correlation of these traits with performances in dressage and jumping competition from offspring of breeding stallions are estimated. Data from 1978-1988 are used, covering scores from 337 3-year-old stallions. Eight subjectively scored traits are considered. These traits are: walk; trot; canter; riding ability; show jumping; free jumping; cross country; character. SPT lasts for a period of 100 days. Data from SPT are analysed using an animal model. The relations between SPT of stallions and performances in jumping and dressage competition are analysed with an animal model for SPT data and a sire model for competition data. Variance and covariance components are estimated by restricted maximum likelihood (REML) procedures. Estimates of heritability are high (0.64) for gaits and riding ability, intermediate (0.41) for cross country and medium-high (0.31) for jumping. Estimated genetic correlation between show jumping scored during SPT and jumping in competition from offspring of breeding stallions is 0.84; for dressage this relation is 0.83. Some possible bias due to selection and the subjectivity of scoring is discussed. It is indicated that selection on SPT of stallions before entering breeding service is an effective tool to breed for ability of performance in competition. |
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refbase @ user @ |
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3962 |
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Huizinga, H.A.; Korver, S.; van der Meij, G.J.W. |
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Stationary performance testing of stallions from the Dutch Warmblood riding horse population. 2. Estimated heritabilities of and correlations between successive judgements of performance traits |
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1991 |
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Livestock Production Science |
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27 |
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2-3 |
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245-254 |
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dressage; genetic parameters; horse; jumping; performance; stallion |
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The length of test period of stationary performance testing (SPT) of stallions of the Dutch Warmblood riding horse population is evaluated. Heritability of successive judgements of traits and the phenotypic and genetic relations between successive judgements are estimated. Data from 1983-1988 are used, covering scores from 206 mostly 3-year-old stallions. Ten subjectively scored traits are considered: walk, trot, canter, riding ability, jumping ability, free jumping, cross country, character, stable behaviour, training report. Traits are successively scored at about 25, 50, 80 and 100 days in SPT. Missing scores are predicted on basis of the available scores using multiple partial regression coefficients. Validity of this method is checked in an independent data set for walk, trot and canter. The correlations between predicted and realized scores average 0.74, 0.77 and 0.79 when first, first and second, and first, second and third judgements are available, respectively. Variance and covariance components are estimated by restricted maximum likelihood (REML) procedures. Data from SPT are analysed using an animal model. Estimates of heritability are high and constant for gaits during the successive judgements. Except for stable behaviour estimates of heritability, the traits decrease slightly during the successive judgements. Estimates of the phenotypic and genetic correlations between successive judgements are high. It is concluded that length of SPT can be shortened and selection during SPT can be intensified. |
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refbase @ user @ |
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3963 |
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