Czerlinski, G. H., Erickson, J. O., & Theorell, H. (1979). Chemical relaxation studies on the horse liver alcohol dehydrogenase system. Physiol Chem Phys, 11(6), 537–569.
Abstract: 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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Domjan, M. (1977). Selective suppression of drinking during a limited period following aversive drug treatment in rats. J Exp Psychol Anim Behav Process, 3(1), 66–76.
Abstract: Administration of lithium chloride disrupted the intake of flavored solutions but not water in rats. This intake suppression was directly related to the amount of lithium administered (Experiment 1), occurred with both palatable and unpalatable novel saccharin solutions (Experiment 2), but was only observed if subjects were tested starting less than 75 min. after lithium treatment (Experiment 3). Twenty-five daily exposures to saccharin did not attenuate the effect (Experiment 4). However, in saccharin-reared and vinegar-reared rats, lithium did not disrupt consumption of the solutions these subjects had access to throughout life, even though suppressions of intake were observed when these subjects were tested with novel flavors (Experiment 5). The selective disruption of drinking is interpreted as a novelty-dependent sensitization reaction to the discomfort of aversive drug administration.
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Mejdell, C. M., Simensen, E., & Boe, K. E. (2005). Is snow a sufficient source of water for horses kept outdoors in winter? A case report. Acta Vet Scand, 46(1-2), 19–22.
Abstract: Due to extreme weather conditions, a flock of outwintered Icelandic horses had to manage for several days on snow as the source of free water. They were fed grass silage ad lib, and any change in feed consumption was not observed. After nine days, blood samples were taken and analysed for plasma osmolality, they were subjected to a simple clinical examination, and offered drinking water. Osmolality levels were within normal limits and mean value did not differ significantly from samples which previously were taken of the same individuals. The general condition of the horses was normal, with no signs of clinical dehydration or disease. The horses showed very little interest for the offered drinking water. This suggests that in cold winter weather, horses being fed grass silage and adjusted to eat snow, can manage for several days with snow substituting liquid water without their physiology and welfare being challenged.
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Rosa, P. A. J., Azevedo, A. M., & Aires-Barros, M. R. (2007). Application of central composite design to the optimisation of aqueous two-phase extraction of human antibodies. J Chromatogr A, 1141(1), 50–60.
Abstract: The partition of human antibodies in aqueous two-phase systems (ATPSs) of polyethylene glycol (PEG) and phosphate was systematically studied using first pure proteins systems and then an artificial mixture of proteins containing 1mg/ml human immunoglobulin G (IgG), 10mg/ml serum albumin and 2mg/ml myoglobin. Preliminary results obtained using pure proteins systems indicated that the PEG molecular weight and concentration, the pH value and the salts concentration had a pronounced effect on the partitioning behaviour of all proteins. For high ionic strengths and pH values higher than the isoelectric point (pI) of the contaminant proteins, IgG could be selectively recovered on the top phase. According to these results, a face centred composite design was performed in order to optimise the purification of IgG from the mixture of proteins. The optimal conditions for the isolation of IgG were observed for high concentrations of NaCl and low concentrations of both phase forming components. The best purification was achieved using an ATPS containing 8% (w/w) PEG 3350, 10% (w/w) phosphate pH 6 and 15% (w/w) NaCl. A recovery yield of 101+/-7%, a purity of 99+/-0% and a yield of native IgG of 97+/-4% were obtained. Back extraction studies of IgG to a new phosphate phase were performed and higher yields were obtained using 10% phosphate buffer at pH 6. The total extraction yield was 76% and the purity 100%.
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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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