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Topic:Biochemistry
The study of biochemistry in horses encompasses the chemical processes and substances that occur within equine organisms. This field investigates the molecular interactions and pathways that are fundamental to horse physiology, including metabolism, enzyme activity, and genetic expression. Key areas of interest include the examination of metabolic disorders, nutrient absorption, and the biochemical basis of muscle function and energy production. Researchers utilize biochemical analysis to understand health and disease mechanisms in horses, contributing to the development of diagnostic tools and therapeutic strategies. This page gathers peer-reviewed studies and scholarly articles that explore various biochemical processes and their implications for equine health and performance.
Thyrotropin stimulation test–new perspective on value of monitoring triiodothyronine. Thyrotropin (thyroid stimulating hormone; TSH) stimulus to thyroid cells of horses and dogs resulted in increased serum triiodothyronine (T3) concentrations that were detected earlier than those of thyroxine (T4). Doubling of the base-line T3 values in horses was detected 0.5 hours after injection of 5 IU of TSH IV, with peak response of 5 times base-line value detected 2 hours after injection. Doubling of T4 values in horses was noticed between 2 and 3 hours, with the peak response of 2.4 times base-line value at 4 hours after injection of TSH. Doubling of base-line T3 values in dogs in respo...
Equine endometrium at pre-implantation stages of pregnancy has specific glycosylated regions. Pre- and post-implantation endometrial tissue was removed from equids at known stages of pregnancy. Thin sections of this material were reacted with fluorescein conjugates of several lectins. Two lectins in particular, with specificities for L-fucose and N-acetylglucosamine, reacted strongly with the endometrial epithelium. The reaction for N-acetylglucosamine occurred in small patches on the endometrial epithelium and extended to cover the entire epithelial surface and endometrial glands after implantation. L-Fucose was observed in larger surface deposits particularly at the openings of endom...
Humoral, hormonal and behavioral correlates of feeding in ponies: the effects of meal frequency. The effect of meal frequency on body fluid, glucose, triiodothyronine (T3), heart rate and behavior was measured in 10 ponies. A simple reversal design was used in which each pony received one meal/day (1X) for 2 wk and six meals/day (6X) for 2 wk. The total intake/day was held constant. Feeding was followed by a rise in plasma levels of glucose, T3, protein and osmolality. One large meal was followed by significantly greater changes in all of the variables than was a meal one-sixth the size. Plasma T3 rose from 41 +/- 5 (SE) ng/liter before feeding to 43 +/- 5 ng/liter following a small meal,...
Hepatobiliary transport of indocyanine green and sulfobromophthalein in fed and fasted horses. Fasting is associated with unconjugated hyperbilirubinemia in several species, including the horse. Studies in ponies showed that a 3-day fast decreased plasma clearance of bilirubin, cholic acid, and sulfobromophthalein (BSP). Since these organic anions are conjugated with different substrates, it is possible that observed differences in plasma clearance result from a general decrease in hepatic conjugating capacity during the animals' fasting. To test this hypothesis, the effects of a 3-day fast on plasma clearance of IV injected BSP (4.4 to 5.1 mg/kg), which is conjugated to glutathione, an...
Effects of multiple intramuscular injections and doses of dexamethasone on plasma cortisol concentrations and adrenal responses to ACTH in horses. Adrenocortical function was assessed in horses given multiple IM doses of dexamethasone to determine the duration of adrenocortical suppression and insufficiency caused by 2 commonly used dosages of dexamethasone (0.044 and 0.088 mg/kg of body weight). Dexamethasone was administered at 5-day intervals for a total of 6 injections. Daily blood samples were collected. The plasma was frozen and later assayed for cortisol. An ACTH response test was determined 2 days before the first injection of dexamethasone and again 8 days after the last dexamethasone injection. Maximum suppression of plasma cor...
Preliminary studies on long distance, retrograde transport of horseradish peroxidase in equine peripheral nerves. As a prelude to studies on retrograde axonal transport of neurotoxin (ie, so-called suicide transport) as a means to prevent post neurectomy neuroma formation, preliminary studies were conducted with an innocuous enzymatic marker, horseradish peroxidase (HRP). The proximal stumps of resected medial and lateral palmar digital nerves in six ponies were injected via a tuberculin syringe and needle with 50 micron 1 of a 30 per cent solution of HRP in order to assess long distance retrograde axonal transport. The dorsal root ganglion of the cervical spinal enlargement (ie, C6, C7, C8, T1, T2) were ...
Gas chromatography-mass spectrometry of androgens in equine ovarian follicles at ultrastructurally defined stages of development. Identification of 19-nortestosterone in follicular fluid. Follicular fluid was obtained from equine follicles at different stages of development as determined by ultrastructural study. Gas chromatography-mass spectrometry associated with stable isotope dilution permitted the demonstration of high levels of 4-estrene-3,7-dione and 17 beta-hydroxy-4-estren-3-one, 17 beta-hydroxy-4-estren-3-one levels often being about 10 times higher than those of testosterone. These findings suggest that in the mare ovary, an aromatizing pathway may proceed using these 19-norsteroids as intermediates. As a consequence of this high level of 19-norsteroids, testosterone...
Fibronectin enhances transfection of Staphylococcus aureus. The factor in normal sera primarily responsible for the enhancement of transfection (and transformation) of Staphylococcus aureus was identified as fibronectin. Serum samples which were depleted of fibronectin by affinity chromatography showed a marked decrease in enhancing activity. Fibronectin isolated from sera of several animal species demonstrated enhancing activity.
Stimulation of equine eosinophil migration by hydroxyacid metabolites of arachidonic acid. Lipoxygenase products of arachidonic acid are important mediators of inflammation, affecting several aspects of cell function. Monohydroxyeicosatetraenoic acid (mono-HETE) and 5,12-dihydroxyeicosatetraenoic acid (LTB4) enhance migration of both neutrophils and eosinophils in several species. The relative ability of positional isomers of HETE and of LTB4 to affect migration of equine eosinophils was studied. The 5, 8, 9, 11, 12, and 15 isomers of HETE were prepared by autooxidation of arachidonic acid, separated by sequential normal phase and reverse phase high performance liquid chromatography...
The time-course of lipid biosynthesis in horse skin. To observe the time-course of formation of sebaceous lipids in the horse, skin was pulse-labelled in vivo by intradermal injection of [1-14C]acetate and the injection sites were harvested at intervals for up to 12 days by skin punch biopsy. The distribution of radioactivity among the major neutral lipid classes and the phospholipids from these biopsies showed that, soon after pulse-labelling, the phospholipids were highly labelled followed by a long-term decrease in radioactivity. Over the same period, the low initial labelling of the dominant component, the equolides (giant ring omega-lactone...
Enhanced biliary bilirubin excretion after heparin-induced erythrocyte mass depletion. The effect of large-dose heparin therapy on erythrocyte mass depletion in ponies was investigated to determine whether stimulation of reticuloendothelial cell activity and catabolic function would be evidenced by enhanced catabolism of heme to bilirubin. Ponies with chronic external biliary fistula were used to examine biliary excretion of bilirubin both before and after heparin loading (107 U/kg, IV, plus 320 U/kg, subcutaneously) and at maintenance dosages of 320 U/kg given (subcutaneously) at 12 and 24 hours after initial loading with heparin. Results indicated that by 48 hours after ponies...
Inhibitory effects of intravenous chloramphenicol sodium succinate on the disposition of phenylbutazone in horses. The effects of i.v. chloramphenicol sodium succinate on the pharmacokinetics of i.v. phenylbutazone in six healthy adult horses were investigated. Administration of chloramphenicol sodium succinate to mares reduced mean (+/- SD) phenylbutazone clearance from 0.600 +/- 0.222 to 0.339 +/- 0.123 ml/min per kg and increased mean (+/- SD) half life from 244 +/- 59.8 to 371 +/- 80.8 min and mean residence time from 333 +/- 86.2 to 533 +/- 124 min. The mean steady-state volume of distribution of phenylbutazone was unchanged, with mean (+/- SD) values of 187 +/- 28.9 ml/kg in control animals and 170 +...
Solubilization and characterization of [3H] 5HT high affinity binding sites (5HT1 and 5HT3). The solubilization of the serotonergic 5HT1 and 5HT3 sites was performed with digitonin and sodium cholate at 1% (final concentration). Two binding sites for [3H]5HT were observed on rat or horse brain synaptosomal membranes solubilized with these detergents. The corresponding dissociation constants (KD) were 1-3 nM and 13-30 nM respectively. These values were closely similar to those corresponding to 5HT1 and 5HT3 sites located in intact membranes. The solubilized sites specifically bound 5HT. The effect of GTP decreasing the binding to 5HT1 sites was lost on solubilized 5HT1 sites; it was re...
Horse kidney neutral alpha-D-glucosidase: purification of the detergent-solubilized enzyme; comparison with the proteinase-solubilized forms. Neutral alpha-D-glucosidase (alpha-D-glucoside glucohydrolase, EC 3.2.1.20) from horse kidney brush-border membranes was solubilized using Emulphogene BC 720 and purified by an affinity chromatography technique. The enzyme preparation (390-fold purified), which was free of other known microvillus hydrolases, exhibited one precipitate line in crossed immunoelectrophoresis and migrated as a single band on sodium dodecyl sulfate polyacrylamide gel electrophoresis. Several criteria (charge-shift crossed immunoelectrophoresis and hydrophobic chromatography) revealed the purified detergent form of t...
Complexities in the denaturation of horse metmyoglobin by guanidine hydrochloride. The denaturation of horse metmyoglobin by guanidine hydrochloride was studied at pH 6.4 and 25 degrees C. Measurements of both the peptide circular dichroism and the absorbance in the Soret region suggest that the extent of renaturation strongly depends on the time interval during which the protein is exposed to concentrated solutions of the denaturant. From the equilibrium measurements of the absorption in the Soret region, it is concluded that the unfolding of metmyoglobin is complex. This is further supported by kinetic studies of denaturation which suggest the occurrence of the least four ...
Activities of key enzymes of aerobic and anaerobic metabolism in middle gluteal muscle from trained and untrained horses. The effect of physical training on the in vitro activities of key enzymes that provide quantitative information on the maximum capacities of anaerobic and aerobic metabolism has been investigated in the gluteal muscle of the horse. Training had no effect on the activities of 6-phosphofructokinase or creatine kinase, suggesting that there was no effect on the capacity of anaerobic metabolism in this muscle. However, the activities of hexokinase and citrate synthase were increased, indicating that training increased the capacity of aerobic metabolism. For comparative purposes, muscle fibre compo...
The isoelectric focusing of keratins in hair followed by silver staining. An isoelectric focusing method followed by silver staining has been developed for the study of keratins which is as effective as two-dimensional electrophoresis and fluorography for hair species identification. Hair from dogs, rabbits, horses, cows, guinea-pigs, donkeys, sheep and cats were successfully identified. Narrow pH ranges were used to observe heterogeneity in human hair. Although this heterogeneity may be affected by environmental conditions, it may be of use in criminalistics.
High resolution R-bands produced in equine chromosomes after incorporation of bromodeoxyuridine. Cell synchronization was used to obtain an adequate percentage of very long chromosomes in equine mitotic spreads. Reported here is our variation, adapted to horse chromosomes, of a method using excess thymidine followed by bromodeoxyuridine incorporation. This technique routinely yields excellent quality cells, predominantly in prometaphase and prophase. Among other differences with the standard technique, this method does not use Colcemid, which, in addition to inhibiting spindle fiber formation, also increases chromosome contraction resulting in thicker and thus fewer bands. Consequently, h...
Equine whole saliva: variability of some major constituents. Whole saliva was collected from six horses over a period of five weeks in sufficient volume for the analysis of 10 constituents. There was considerable variation in the concentration of the analytes both between horses and between different days in the same horse. The most variable constituent was sodium, and the least variable was glucose, but this was derived from the sweet used to stimulate salivation. The use of whole saliva as a fluid for investigation would depend on achieving the minimum variability possible.
Disposition of sulfadimidine and its N4-acetyl and hydroxy metabolites in horse plasma. The plasma disposition of sulfadimidine (SDM) and its metabolites N4-acetylsulfadimidine (N4-SDM), 6-hydroxymethyl-4-methyl-pyrimidine (SCH2OH) and 5-hydroxy-4,6-dimethyl-pyrimidine (SOH), was studied in three horses following intravenous administration of SDM at dose levels of 20 and 200 mg/kg in cross-over trials. The percentages of N4-SDM (0.58-0.90%), SOH (0.83-6.75%) and SCH2OH (0.38-0.71%) in plasma, expressed as a percentage of the total sulfonamide concentration, were small and their plasma concentrations were parallel with SDM from 4 h following administration. At high doses (200 mg/k...
Incorporation of L-75Se-cystine in tissue fragments from the matrix of the hoof and the claw–a tool for studying the pathogenesis of laminitis? An in vitro method has been designed and used to study the incorporation of 75Se-cystine into matrix fragments from hooves and claws of healthy horses and cattle. Tissue fragments from the zone of keratinisation were incubated with L-75Se-cystine in a tissue culture medium for 4 to 6 h, during which time there was continuous incorporation of the labelled selenocystine. The incorporation was greatly decreased by adding L-cystine to the incubation mixture. It is concluded that the incorporation of 75Se-cystine depends on the presence of a specific receptor for cystine in the tissue fragments stu...
The influence of dietary selenium levels on blood levels of selenium and glutathione peroxidase activity in the horse. Twenty mature geldings, averaging 535 kg, were used to determine the influence of dietary selenium (Se) on the blood levels of Se and Se-dependent glutathione peroxidase (SeGSH-Px) activity in the horse. Horses were randomly assigned within breed to four treatments consisting of five horses each and fed a basal diet containing .06 ppm of naturally occurring Se. Diets were supplemented with .05, .10 and .20 ppm Se, as sodium selenite. Blood was drawn for 2 wk before, and for 12 wk following, the inclusion of supplement Se in the diets. Whole blood and plasma Se concentrations and plasma SeGSH-P...
Properties of monoclonal antibodies against glycoproteins of western equine encephalitis virus. To analyze the biological activities of the alphavirus glycoproteins, eight different monoclonal antibodies against the two glycoproteins of western equine encephalitis virus were isolated. Five of the eight monoclonal antibodies were shown to be specific for E1 and three for E2 protein by an enzyme-linked immunosorbent assay and by radioimmunoprecipitation. Three of the five anti-E1 and all of the anti-E2 monoclonal antibodies inhibited hemagglutination by purified virions. One anti-E1 and two anti-E2 monoclonal antibodies possessed high virus-neutralizing activity.
Demonstration of a COOH-terminal extension on equine lutropin by means of a common acid-labile bond in equine lutropin and equine chorionic gonadotropin. The beta subunits of equine lutropin and equine chorionic gonadotropin were incubated in 0.013 N HCl for 30 min at 110 degrees C and separated into two fragments by reverse-phase high performance liquid chromatography. The amino acid and carbohydrate compositions of both fragments from each subunit were analyzed. The results demonstrated that equine lutropin-beta has a glycosylated COOH-terminal extension that differs only in carbohydrate composition from the COOH-terminal portion of equine chorionic gonadotropin-beta. This is the first demonstration of a glycosylated COOH-terminal extension i...
Kinetic studies of the unfolding-refolding of horse muscle phosphoglycerate kinase induced by guanidine hydrochloride. The kinetics of the unfolding and refolding of horse muscle phosphoglycerate kinase were studied with three different signals: fluorescence emission intensity at 336 nm (excitation at 292 nm), ellipticity at 220 nm, and enzyme activity. The results corroborate the conclusion on the existence of intermediates in the folding pathway obtained from equilibrium studies. Kinetic studies showed at least two phases of refolding, as revealed by fluorescence as well as by circular dichroism measurements. During the fast phase, an intermediate was formed with a fluorescence intensity higher than that of ...
Mechanism of renal excretion of creatinine by the pony. Free-flow and stop-flow procedures conducted on 2 female and 2 testosterone-treated castrated male ponies indicated that [14C]inulin and exogenous creatinine clearance values were the same. These results indicated that creatinine was neither reabsorbed nor secreted by the renal tubules and that exogenous creatinine clearance was an accurate method for determining glomerular filtration rate. As in other species which have been studied, endogenous creatinine clearance probably underestimated glomerular filtration rate because of the presence of noncreatinine chromogens in plasma.
Proteinase inhibitors of horse seminal plasma. A high molecular mass, acid-soluble proteinase inhibitor. Horse seminal plasma does not possess a proteinase inhibitor corresponding to human HUSI-I (human seminal plasma inhibitor). Instead a protein complex of high relative molecular mass (Mr) containing proteinase inhibitory activity was detected, which was called horse seminal plasma protein complex or HSPC. The compound had a broad enzyme-inhibiting spectrum. Its Mr was estimated to be 800 000 and it was composed of 7 different polypeptides with Mr values ranging from 11 000 to 30 000. Its carbohydrate content was between 3.5% and 5%. Despite the high molecular mass, the complex was soluble in d...
Pharmacokinetics of small doses of 3-methylindole given to horses. The pharmacokinetics of 3-methylindole (3MI) given orally in 2 doses (10 mg/kg and 20 mg/kg) to horses were determined. The pharmacokinetic plasma-concentration profiles for 3MI (10- and 20-mg/kg dosages) in horses were represented by a 2-compartment open model with first-order absorption, as determined by nonlinear least-squares regression analysis. Absorption of 3MI at both dosages was rapid. Comparisons of the peak plasma concentrations, the postdistribution half lives, total clearances, and areas under the curve of the plasma-concentration profiles between the 10- and the 20-mg/kg dosages ...
