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Shaw, E. B., Houpt, K. A., & Holmes, D. F. (1988). Body temperature and behaviour of mares during the last two weeks of pregnancy. Equine Vet J, 20(3), 199–202.
Abstract: Average daily core body temperature and behavioural patterns of pregnant mares were studied, in search of definitive signs of parturition within 24 h of the event. Nineteen pony mares were sampled twice daily for core body temperature. A significant temperature drop, averaging 0.1 degrees C (0.2 degrees F) was observed during the day prior to parturition. Between 18.00 h and 06.00 h, during the two weeks before parturition, Thoroughbred and Standardbred mares (n = 52) spent an average 66.8 per cent of their time standing, 27.0 per cent eating, 4.9 per cent lying in sternal recumbency, 1.0 per cent lying in lateral recumbency, and 0.3 per cent walking. On the night before parturition, mares spent significantly less time lying in sternal recumbency than on previous nights and on the night of parturition all behaviour patterns except eating were significantly different from the nights of the two weeks before parturition. There was an increase in walking (5.3 per cent), lying in sternal recumbency (8 per cent) and lying in lateral recumbency (5.3 per cent) whereas standing (53.3 per cent) was decreased. In 58 observed pregnancies, 54 mares (97 per cent) foaled in a recumbent position and 50 mares (86 per cent) foaled between 18.00 h and 06.00 h.
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Steinhoff, H. J., Lieutenant, K., & Redhardt, A. (1989). Conformational transition of aquomethemoglobin: intramolecular histidine E7 binding reaction to the heme iron in the temperature range between 220 K and 295 K as seen by EPR and temperature-jump measurements. Biochim Biophys Acta, 996(1-2), 49–56.
Abstract: Temperature-dependent EPR and temperature-jump measurements have been carried out, in order to examine the high-spin to low-spin transition of aquomethemogobin (pH 6.0). Relaxation rates and equilibrium constants could be determined as a function of temperature. As a reaction mechanism for the high-spin to low-spin transition, the binding of N epsilon of His E7 to the heme iron had been proposed; the same mechanism had been suggested for the ms-effect, found in temperature-jump experiments on aquomethemoglobin. A comparison of the thermodynamic quantities, deduced form the measurements in this paper, gives evidence that indeed the same reaction is investigated in both cases. Our results and most of the findings of earlier studies on the spin-state transitions of aquomethemoglobin, using susceptibility, optical, or EPR measurements, can be explained by the transition of methemoglobin with H2O as ligand (with high-spin state at all temperatures) and methemoglobin with ligand N epsilon of His E7 (with a low-spin ground state). Thermal fluctuations of large amplitude have to be postulated for the reaction to take place, so this reaction may be understood as a probe for the study of protein dynamics.
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Dunn, M. F., & Branlant, G. (1975). Roles of zinc ion and reduced coenzyme in horse liver alcohol dehydrogenase catalysis. The mechanism of aldehyde activation. Biochemistry, 14(14), 3176–3182.
Abstract: 1,4,5,6-Tetrahydronicotinamide adenine dinucleotide (H2NADH) has been investigated as a reduced coenzyme analog in the reaction between trans-4-N,N-dimethylaminocinnamaldehyde (I) (lambdamax 398 nm, epsilonmax 3.15 X 10-4 M-minus 1 cm-minus 1) and the horse liver alcohol dehydrogenase-NADH complex. These equilibrium binding and temperature-jump kinetic studies establish the following. (i) Substitution of H2NADH for NADH limits reaction to the reversible formation of a new chromophoric species, lambdamax 468 nm, epsilonmax 5.8 x 10-4 M-minus 1 cm-minus 1. This chromophore is demonstrated to be structurally analogous to the transient intermediate formed during the reaction of I with the enzyme-NADH complex [Dunn, M. F., and Hutchison, J. S. (1973), Biochemistry 12, 4882]. (ii) The process of intermediate formation with the enzyme-NADH complex is independent of pH over the range 6.13-10.54. Although studies were limited to the pH range 5.98-8.72, a similar pH independence appears to hold for the H2NADH system. (iii) Within the ternary complex, I is bound within van der Waal's contact distance of the coenzyme nicotinamide ring. (iv) Formation of the transient intermediate does not involve covalent modification of coenzyme. Based on these findings, we conclude that zinc ion has a Lewis acid function in facilitating the chemical activation of the aldehyde carbonyl for reduction, and that reduced coenzyme plays a noncovalent effector role in this substrate activating step.
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Wilson, M. T., Ranson, R. J., Masiakowski, P., Czarnecka, E., & Brunori, M. (1977). A kinetic study of the pH-dependent properties of the ferric undecapeptide of cytochrome c (microperoxidase). Eur J Biochem, 77(1), 193–199.
Abstract: The ferric form of the haem undecapeptide, derived from horse cytochrome c by peptic digestion, undergoes at least three pH-induced transitions with pK values of 3.4, 5.8 and 7.6. Temperature-jump experiments suggest that the first of these is due to the binding of a deprotonated imidazole group to the feric iron while the second and third arise from the binding of the two available amino groups present (the alpha-NH2 of valine and the epsilon-NH2 of lysine). Molecular models indicate that steric retraints on the peptide dictate that these amino groups may only coordinate to iron atoms via intermolecular bonds, thus leading to the polymerization of the peptide. Cyanide binding studies are in agreement with these conclusions and also yield a value of 3.6 X 10(6) M-1 s-1 for the intrinsic combination constant of CN- anion with the haem. A model is proposed which describes the pH-dependent properties of the ferric undecapeptide.
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Kihara, H., Nakatani, H., Hiromi, K., & Hon-Nami, K. (1977). Kinetic studies on redox reactions of hemoproteins. I. Reduction of thermoresistant cytochrome c-552 and horse heart cytochrome c by ferrocyanide. Biochim Biophys Acta, 460(3), 480–489.
Abstract: The oxidation-reduction reaction of horse heart cytochrome c and cytochrome c (552, Thermus thermophilus), which is highly thermoresistant, was studied by temperature-jump method. Ferrohexacyanide was used as reductant. (Formula: see text.) Thermodynamic and activation parameters of the reaction obtained for both cytochromes were compared with each other. The results of this showed that (1) the redox potential of cytochrome c-552, + 0.19 V, is markedly less than that of horse heart cytochrome c. (2) deltaHox of cytochrome c-552 is considerably lower than that of horse heart cytochrome c. (3) deltaSox and deltaSred of cytochrome c-552 are more negative than those of horse heart cytochrome c. (4) kred of cytochrome c-552 is much lower than that of horse heart cytochrome c at room temperature.
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Arnold, W., Ruf, T., & Kuntz, R. (2006). Seasonal adjustment of energy budget in a large wild mammal, the Przewalski horse (Equus ferus przewalskii) II. Energy expenditure. J Exp Biol, 209(Pt 22), 4566–4573.
Abstract: Many large mammals show pronounced seasonal fluctuations of metabolic rate (MR). It has been argued, based on studies in ruminants, that this variation merely results from different levels of locomotor activity (LA), and heat increment of feeding (HI). However, a recent study in red deer (Cervus elaphus) identified a previously unknown mechanism in ungulates--nocturnal hypometabolism--that contributed significantly to reduced energy expenditure, mainly during late winter. The relative contribution of these different mechanisms to seasonal adjustments of MR is still unknown, however. Therefore, in the study presented here we quantified for the first time the independent contribution of thermoregulation, LA and HI to heart rate (f(H)) as a measure of MR in a free-roaming large ungulate, the Przewalski horse or Takhi (Equus ferus przewalskii Poljakow). f(H) varied periodically throughout the year with a twofold increase from a mean of 44 beats min(-1) during December and January to a spring peak of 89 beats min(-1) at the beginning of May. LA increased from 23% per day during December and January to a mean level of 53% per day during May, and declined again thereafter. Daily mean subcutaneous body temperature (T(s)) declined continuously during winter and reached a nadir at the beginning of April (annual range was 5.8 degrees C), well after the annual low of air temperature and LA. Lower T(s) during winter contributed considerably to the reduction in f(H). In addition to thermoregulation, f(H) was affected by reproduction, LA, HI and unexplained seasonal variation, presumably reflecting to some degree changes in organ mass. The observed phase relations of seasonal changes indicate that energy expenditure was not a consequence of energy uptake but is under endogenous control, preparing the organism well in advance of seasonal energetic demands.
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Permyakov, S. E., Khokhlova, T. I., Nazipova, A. A., Zhadan, A. P., Morozova-Roche, L. A., & Permyakov, E. A. (2006). Calcium-binding and temperature induced transitions in equine lysozyme: new insights from the pCa-temperature “phase diagrams”. Proteins, 65(4), 984–998.
Abstract: The most universal approach to the studies of metal binding properties of single-site metal binding proteins, i.e., construction of a “phase diagram” in coordinates of free metal ion concentration-temperature, has been applied to equine lysozyme (EQL). EQL has one relatively strong calcium binding site and shows two thermal transitions, but only one of them is Ca(2+)-dependent. It has been found that the Ca(2+)-dependent behavior of the low temperature thermal transition (I) of EQL can be adequately described based upon the simplest four-states scheme of metal- and temperature-induced structural changes in a protein. All thermodynamic parameters of this scheme were determined experimentally and used for construction of the EQL phase diagram in the pCa-temperature space. Comparison of the phase diagram with that for alpha-lactalbumin (alpha-LA), a close homologue of lysozyme, allows visualization of the differences in thermodynamic behavior of the two proteins. The thermal stability of apo-EQL (transition I) closely resembles that for apo-alpha-LA (mid-temperature 25 degrees C), while the thermal stabilities of their Ca(2+)-bound forms are almost indistinguishable. The native state of EQL has three orders of magnitude lower affinity for Ca(2+) in comparison with alpha-LA while its thermally unfolded state (after the I transition) has about one order lower (K = 15M(-1)) affinity for calcium. Circular dichroism studies of the apo-lysozyme state after the first thermal transition show that it shares common features with the molten globule state of alpha-LA.
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Pritchard, J. C., Barr, A. R. S., & Whay, H. R. (2006). Validity of a behavioural measure of heat stress and a skin tent test for dehydration in working horses and donkeys (Vol. 38).
Abstract: REASONS FOR PERFORMING STUDY: Dehydration and heat stress are serious welfare issues for equids working in developing countries. There is a lack of any standardised method or validated interpretation of the skin tent test in horses and donkeys. Owners of dehydrated and heat-stressed animals often depend on veterinary examination for identification of these conditions, leading to delays in treatment and unnecessary reliance on external sources to effect welfare improvement. OBJECTIVES: To validate a standardised skin tent test for dehydration and a behavioural measure of heat stress in working equids; and to examine the effect of heat stress and dehydration on tripping and staggering behaviour. METHODS: The study was carried out on 130 working horses and donkeys in Pakistan. Associations between skin tent and blood parameters (packed cell volume [PCV], serum total protein [TP], serum osmolality), clinical parameters, resting and drinking behaviour were examined. Heat stress behaviour (increased respiratory rate and depth, head nodding, flared nostrils, apathy) was observed in conjunction with rectal temperature. Tripping and staggering were assessed using a simple obstacle course. RESULTS: In both species, heat stress behaviour was significantly associated with increased rectal temperature (P<0.001). A positive skin tent test was not significantly associated with PCV or TP, although in donkeys it was significantly associated with lower serum osmolality (P<0.001). More animals age >15 years had a positive skin tent than those in younger age groups (P = 0.037). Very thin horses were more likely to have a positive skin tent than those in thin or moderate condition (P = 0.028). There was no significant correlation between skin tent and tripping or staggering in either species. CONCLUSIONS AND POTENTIAL RELEVANCE: Heat stress behaviour is related to increased body temperature in working horses and donkeys. Owners may use this to make judgements regarding rest and cooling, precluding the need to seek veterinary attention. The skin tent test for dehydration used in this study did not show a significant relationship with PCV or TP. However, the use of blood parameters to validate the skin tent test may be confounded by anaemia, hypoproteinaemia or electrolyte depletion. Alternative methods are needed to confirm or refute the validity of the skin tent test in working equids.
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Balakrishnan, G., Hu, Y., & Spiro, T. G. (2006). Temperature-jump apparatus with Raman detection based on a solid-state tunable (1.80-2.05 microm) kHz optical parametric oscillator laser. Appl Spectrosc, 60(4), 347–351.
Abstract: 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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Piccione, G., Caola, G., & Refinetti, R. (2005). Temporal relationships of 21 physiological variables in horse and sheep. Comp Biochem Physiol A Mol Integr Physiol, 142(4), 389–396.
Abstract: Daily or circadian oscillation has been documented in a variety of physiological and behavioral processes. Although individual variables have been studied in great detail, very few studies have been conducted on the temporal relationships between the rhythms of different variables. It is not known whether the circadian pacemaker generates each and every rhythm individually or whether most rhythms are simply derived from a few clock-controlled rhythms. As a first step in elucidating this issue, 21 physiological variables were recorded simultaneously in horse and sheep. The results indicated that, in both species, different variables exhibit different degrees of daily rhythmicity and reach their daily peaks at different times of the day. The variables exhibiting strongest rhythmicity were locomotor activity, rectal temperature, and plasma concentrations of melatonin and glucose. Comparison of rhythmicity and acrophase in the various rhythms allowed inferences to be made about mechanisms of causation.
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