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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.
Effects of exercise on plasma concentrations of caffeine and its metabolites in horses. The effects of exercise on the metabolism of caffeine (CA) were studied 3h after administration of the drug to race horses which then underwent exercise sets (1000-m gallop). Analysis was made of pharmacokinetics of CA, changes in its plasma concentrations, its metabolites, i.e., theophylline (TP), theobromine (TB) and paraxanthine (PX), and the molar concentration ratios of CA to these metabolites. After exercise, AUC and t1/2 tended to decrease, and the concentration of CA decreased, while the concentrations of TP and TB significantly increased. The TP/CA ratio and TB/CA ratio significantly ...
Blood ionized calcium concentrations in horses before and after the cross-country phase of three-day event competition. Blood ionized calcium (Ca2+) and pH; plasma lactate concentrations; and total protein, total calcium (CaT), albumin, and phosphorus concentrations in serum were determined in 40 healthy horses before (T1), at the finish line (T2), and 10 minutes after the finish (T3) of the cross-country phase of a 3-day-event competition. Mean (+/- SEM) Ca2+ concentrations decreased from 6.22 +/- 0.04 mg/dl at T1 to 5.04 +/- 0.07 mg/dl at T2 (P < or = 0.05). This decrease was accompanied by a nonsignificant increase in CaT between T1 and T2. The mean (+/- SEM) percent ionization of calcium decreased signif...
Equine sweat composition: effects of adrenaline infusion, exercise and training. Significant alterations in plasma electrolyte concentrations have been reported in horses following prolonged exercise, resulting from loss of hypertonic sweat. Sweat was collected from 10 horses undergoing a 10 week training programme; 5 at moderate intensity, to speeds of 10 m/s and 5 at low intensity, to speeds of 5 m/s. Sweat was collected from 2 sites in response to a submaximal exercise test (30 min at 50% VO2max and during an adrenaline infusion (dose mean +/- s.d.; 0.3 +/- 0.05 g/kg over 30 min). Sweat samples were analysed for sodium, chloride, potassium, protein, magnesium, calcium a...
Demonstration of tissue-specific promoters in nonprimate species that express aromatase P450 in placentae. Conversion of androgens to estrogens is catalyzed by aromatase P450 (P450arom; the product of the CYP19 gene). Regulation of tissue-specific expression of P450arom in humans is due, in part, to alternative transcriptional start sites that arise as a consequence of the use of granulosa cells and placental tissue from cows, horses, and pigs (ungulates) in order to determine whether these species, like the human, utilize tissue-specific promoters to drive P450arom expression. The majority of transcripts in the placenta have 5'-termini that differ from those in the ovary upstream of a common site ...
Cryopreservation reduces the ability of equine spermatozoa to attach to oviductal epithelial cells and zonae pellucidae in vitro. Two bioassays were used to evaluate the interaction of fresh and cryopreserved equine semen with oviductal epithelial cells (OEC) and with the zona pellucida (ZP). Split ejaculates were either stored at room temperature or frozen and thawed. In experiment 1, progressive motility and membrane integrity were evaluated for each treatment. Fluorescent labeled spermatozoa were cocultured with monolayers of OEC for 30 minutes, and the number of sperm attached to OEC was counted by fluorescence microscopy and analysis of digitized images. Motility of spermatozoa attached to OEC was observed at 0.5, 3...
Causes of differences in respiration rate of hepatocytes from mammals of different body mass. Resting O2 consumption of hepatocytes isolated from mammals ranging in mass from 20-g mice to 200-kg horses decreases with increasing body mass. The substrate oxidation system increases in activity with increasing body mass and mitochondrial proton leak and phosphorylation system decrease in activity, resulting in a higher mitochondrial membrane potential in hepatocytes from larger mammals. The absolute rates of O2 consumption due to nonmitochondrial processes, substrate oxidation, mitochondrial proton leak, and the phosphorylation system decrease with increasing body mass. These decreases are...
Effect of hepatic isoferritins from iron overloaded rats on lymphocyte proliferative response: role of ferritin iron content. Iron and ferritin impair a variety of immunological functions. To evaluate the effect of ferritin iron content on rat lymphocyte proliferative response, isoferritins that differ in their iron content and isoelectric point (pI) were isolated from iron overload rat livers by ultracentrifugation (isoferritins with high iron content and low pI) or crystallization (isoferritins with low iron content and high pI) methods. Additionally, commercial horse splenic ferritin (with a lower pI and higher iron content than rat isoferritins) was also tested. Proliferative response to Con A was decreased in a ...
Effects of shortening the steeplechase phase (phase B) of a 3-day-event. Thirty-four horses competing in the Endurance Test of a 3-day-event were divided into 3 groups: horses in Group 1 (n = 15) competing in a 3.5 min steeplechase phase; horses in Group 2 (n = 13) in a 3 min steeplechase phase (Phase B) and horses in Group 3 (n = 6) in a 2.5 min steeplechase phase. The shortening of Phase B was associated with a lengthening of Phase C so that the total distance of the event for all horses was 14,940 m. Bodyweight (BW) was measured and total body water (TBW) and water loss estimated. Blood samples were collected from the horses prior to the Endurance Test, at the e...
Plasma volume and ions during exercise in cool, dry; hot, dry; and hot, humid conditions. We studied the effects of heat and relative humidity (RH) on plasma volume (PV) and ion responses to submaximal exercise and 60 min recovery in Thoroughbreds. Five horses were exercised at 50% of peak VO2 in cool, dry (CD, T = 22 degrees C, RH = 45-55%), and hot, humid (HH, T = 30-34 degrees C, RH = 80-85%) conditions until a pulmonary artery temperature of 41.5 degrees C was reached. Blood was obtained from the carotid artery. Body mass was measured at rest and after 30 min of recovery. The thermal conditions had no effect on the PV and ion responses during exercise and initial 30 min of reco...
Variable-temperature study of the heme-reorientation process in equine myoglobin. The redistribution of the initially-formed myoglobin heme-insertion isomers from the initially formed 50/50 mixture to the equilibrium ratio of 90/10 has long been assumed to occur by one of two mechanisms, both of which require the rupture of the heme iron-protein bond (La Mar, G.N., Toi, H. and Krishnamoorthi, K. (1984) J. Am. Chem. Soc. 106, 6395-6401). In this study we compared the use of nuclear magnetic resonance and optical spectroscopic techniques as methods for studying the reorientation of heme within myoglobin. We found that kinetics determinations of the heme insertion isomer redis...
Metalloproteinase activity has a role in equine chorionic girdle cell invasion. Chorionic girdle cells are a highly invasive subpopulation of trophoblast cells of the equine conceptus. By Day 35 (Day 0 = day of ovulation), cells of the chorionic girdle adhere to the uterine epithelium and begin to invade the endometrial wall. Invasive cells must attach to extracellular matrix proteins, secrete proteinases capable of degrading matrix, and migrate through the degraded matrix; invasion is largely dependent on the proteinase activity of the cells. The objective, therefore, was to develop an in vitro system to examine the mechanisms of equine chorionic girdle cell invasion thr...
Effect of tumor necrosis factor antibody on synovial fluid cytokine activities in equine antebrachiocarpal joints injected with endotoxin. Six horses received intra-articular injections of a mixture of 1 micrograms of endotoxin/5 mg of equine tumor necrosis factor (eqTNF) monoclonal antibody in 1 antebrachiocarpal joint and an equal volume (2 ml) of 1 micrograms of endotoxin/5 mg of control antibody in the opposite joint. Synovial fluid sample collection (1 ml) was accomplished by use of an indwelling, intra-articular catheter at postinjection hours (PIH) 0, 1, 1.5, 2, 5, and 8, and by arthrocentesis at PIH 24. Joint fluid samples were analyzed for nucleated cell count, protein concentration, and TNF, interleukin 6 (IL-6), IL-1, ...
Simultaneous analysis of tiaramide metabolites in horse urine and plasma by solid-phase extraction and reversed-phase ion-pair liquid chromatography. A simple method for the simultaneous analysis of tiaramide (TRA) metabolites in the horse is described. The sample preparation method using a Bond-Elut PH cartridge and stepwise elution with ice-cold, 30% aqueous methanol followed by additional methanol is effective for recovering the metabolites with different properties. The extraction method gives good recoveries (greater than 80%) and reproducibility. Each metabolite is well separated by high-performance liquid chromatography using an octadecyl-type column of polymer-based packing with a solvent system of 20 mM phosphate buffer (pH 6.5)-ac...
Sandwich enzyme-linked immunosorbent assay for quantitative measurement of serum amyloid A protein in horses. To measure the concentration of serum amyloid A (sAA) protein in horses, a sensitive and highly reproducible sandwich (ELISA) was established, using affinity purified SAA antibody. Results of the ELISA were found to have a high correlation (r = 0.95) with those of the single radial immunodiffusion test. Equine SAA concentration was measured by use of this ELISA. In clinically normal horses, the concentration of SAA was high immediately after birth to 2 weeks of age. After that, SAA concentration had periodic fluctuations in the range of approximately 1.0 to 30 micrograms/ml. Mean (+/- SD)) con...
Effect of different protein supplements on motility and plasma membrane integrity of frozen-thawed stallion spermatozoa. Three experiments were conducted to evaluate the effect of different macromolecule components (egg yolk, skim milk, and BSA) in a widely employed extender for cryopreservation of horse semen. Spermatozoal motility (MOT) and the percentage of spermatozoa with an intact plasma membrane (IPM) were evaluated in frozen-thawed samples. In the first experiment (four Draft Horse stallions, four ejaculates each) a standard freezing extender containing 20% whole egg yolk was modified by replacing extender components (glucose-EDTA solution, 11% lactose solution) with an increasing volume of a skim milk d...
[Adenosine deaminase activity in blood and tissues of horses of the Rassen Haflinger and Thüringer Kaltblut breeds]. The activity of adenosine deaminase in the cerebrum, cerebellum, brain stem, heart, skeletal muscle, lung, liver, spleen, kidney and testes of horses of the breeds Haflinger and Thüringer Kaltblut was not different from each other. In the erythrocytes and the plasma no activity of the ADA exists.
The unfolding thermodynamics of c-type lysozymes: a calorimetric study of the heat denaturation of equine lysozyme. The energetics of the temperature-induced unfolding of equine lysozyme was studied calorimetrically and compared with that of two structurally homologous proteins: hen egg white lysozyme and alpha-lactalbumin. The structure of each of these proteins is characterized by the presence of a deep cleft that divides the molecule into two regions called the alpha and beta domains. In equine lysozyme and alpha-lactalbumin the latter domain specifically binds Ca2+. It is shown that, in contrast to hen egg white lysozyme in which the alpha and beta domains unfold as a single cooperative unit, in equine ...
Beta-thiopropionyl cytochromes c modified at lysyl residues: preparation and characterization of the monosubstituted horse cytochromes c. beta-Thiopropionyl derivatives of horse cytochrome c singly modified at each of 18 different lysine epsilon-amino groups have been prepared using sulfosuccinimidyl-2-(biotinamido)ethyl-1,3-dithiopropionate and purified to homogeneity by high-pressure liquid chromatography. These derivatives were characterized by determination of: (i) the location of the modification; (ii) reduction potentials; (iii) visible and NMR spectra: and by (iv) measurement of electron transfer activity with cytochrome-c oxidase. No significant changes in structure were indicated, except for the ferric forms of the deri...
Effect of glycosylation on the heparin-binding capability of boar and stallion seminal plasma proteins. Boar and stallion seminal plasmas were fractionated using affinity chromatography on heparin-Sepharose. In both species, among other proteins, the heparin-binding (H+) and non-heparin-binding (H-) fractions each contained glycoforms of either porcine PSP-I or equine HSP-1 and HSP-2. However, porcine H+/PSP-I eluted as a monomeric protein, whereas H-/PSP-I formed a heterodimer with PSP-II, another major seminal plasma protein. On the other hand, the stallion proteins H+/HSP-1 and H+/HSP-2 eluted together as an aggregate of relative molecular mass (M(r)) 90,000, whereas H-/HSP-1 and H-/HSP-2 elu...
Effect of glucocorticoids on serum osteocalcin concentration in horses. The effects of dexamethasone (0.2 mg/kg of body weight; IV, IM, and PO) and methylprednisolone acetate (120 mg given intra-articularly) on serum osteocalcin and cortisol concentrations were studied in 6 horses. Serum osteocalcin and cortisol concentrations were serially monitored after each treatment. A significant (P < 0.05) decrease in serum osteocalcin and cortisol concentrations was observed from 12 to 24 and 2 to 48 hours, respectively, after IV and IM administrations of dexamethasone. Serum osteocalcin and cortisol concentrations were significantly decreased from 6 to 48 and 3 to 72 h...
Modulation of 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) activity in the equine placenta by pregnenolone and progesterone metabolites. The purpose of this study was to measure 3beta-HSD activity in the equine placenta and to assess the effect of fetal and maternal blood plasma progestagens on 3beta-HSD activity was measured in 8 late gestation (collected by caesarian section at 250 to 320 days) and 7 term (collected by caesarian section at 250 to 320 days) and 7 term (collected at birth) equine placentae using a tritium release assay with [3alpha-3H] pregnenolone as substrate. Mean +/- s.d. Km(app) and Vmax for term placentae were in general higher than for late gestation placentae (0.129 +/- 0.217 micromol/l and 23.85 +/- 9....
In vitro Catabolism of very low density lipoproteins from horse (Equus caballus) by the action of autologous lipoprotein lipase. Incubation of equine very low density lipoproteins with lipoprotein lipase isolated from horse postheparin plasma resulted in the formation of lipoproteins of a higher density. Lipoproteins isolated after incubation and plasma lipoproteins had a different chemical composition and triacylglycerol fatty acid pattern. In vitro-obtained low density lipoproteins contained substantially more phospholipids and triacylglycerols but significantly less cholesteryl esters than native low density lipoproteins. Comparing the triacylglycerol fatty acid pattern of plasma very low density lipoproteins and in ...
Quinidine administration increases steady state serum digoxin concentration in horses. The aim of this study was to determine if quinidine administration increases steady state serum digoxin concentration in horses. Digoxin (0.01 mg/kg q. 12 h per os) was administered to 6 horses for 7 days. Steady state was confirmed by identifying statistically indistinguishable peak and trough serum digoxin concentrations on Days 4, 5, and 6. On Day 6, serum digoxin concentration was measured at baseline and 0.25, 0.5, 1, 2, 4, 6, 8 and 12 h after digoxin administration. On Day 7, quinidine (20 g at baseline and 10 g at 2, 4 and 6 h) was administered per os and serum digoxin concentration was...
Localisation of 15-hydroxy prostaglandin dehydrogenase (PGDH) and steroidogenic enzymes in the equine placenta. 15-hydroxy prostaglandin dehydrogenase (PGDH) is the critical enzyme that determines metabolism of primary prostaglandins. Its expression is determined in part by steroid hormones, particularly progesterone, formed from delta(5) steroids through 3beta-hydroxysteroid dehydrogenase (3beta-HSD) activity. To assess whether the regulation of PGDH might occur in a paracrine, autocrine or intracrine fashion, we used immunohistochemistry (IHC) to determine the localisation of key steroidogenic enzymes in the equine placenta and compared these patterns to the distribution of immunoreactive (IR-) PGDH. ...
The response of the skeleton to physical training: a biochemical study in horses. In this study we tested the hypothesis that exercise induces an adaptive response in the developing skeleton which may be monitored in vivo by measuring biochemical markers of bone metabolism. The effects of exercise on two biochemical markers of bone formation were determined; the carboxy-terminal propeptide of type I procollagen (PICP), and the bone-specific isoenzyme of alkaline phosphatase (BAP), and one putative marker of resorption, the pyridinoline crosslinked telopeptide domain of type I collagen (ICTP). All three markers were measured for a year in 2-year-old thoroughbred horses exerc...
Effects of sodium bicarbonate on fluid, electrolyte and acid-base balance in racehorses. Sodium bicarbonate given by nasogastric tube has been used by some trainers as the key ingredient in a 'milkshake'. It has been suggested that such treatment given 3-5 h prior to racing may enhance a horse's racing performance by increasing the blood buffering capacity and enhancing lactate clearance from skeletal muscle, thereby delaying the onset of fatigue. Several experiments were conducted to examine the effects on fluid, electrolyte and acid-base values of 0.5 g kg-1 dose of sodium bicarbonate, were examined. The effects of fasting, the simultaneous administration of glucose (0.5 g kg-1)...
Characterisation of beta-adrenoceptors in equine digital veins: implications of the modes of vasodilatory action of isoxsuprine. Isolated equine digital veins (EDVs) were used to study beta-adrenoceptor mediated vasodilation and to examine isoxsuprine's vasodilatory mechanism of action. When the blood vessel wall tension was raised with potassium chloride solution (KCl; 59 mmol/l), the order of vasodilator potency of beta-agonists was: isoprenaline > fenoterol > noradrenaline > dobutamine > isoxsuprine. The beta 2-selective adrenoceptor antagonist, ICI 118551 (1 nmol/l) caused a 6.74 and 6.65-fold parallel shift to the right in the dose response curves to fenoterol and noradrenaline respectively. Propranolol...
A novel uterine protein that associates with the embryonic capsule in equids. An apparently unique protein produced in large quantities by the endometrium of the mare which adheres to, or is incorporated into, the acellular capsule that surrounds the equine conceptus in early pregnancy, has been characterized and partially sequenced. It has a molecular mass of approximately 18 kDa on SDS-PAGE gels and is nonglycosylated as assessed by a sensitive carbohydrate detection kit. Comparison of its first 24 amino-terminal amino acids with all entries in the databases failed to show any significant identity with any other protein sequence. Secretion of the protein appears to be...
Amino acid sequence of HSP-1, a major protein of stallion seminal plasma: effect of glycosylation on its heparin- and gelatin-binding capabilities. We report the complete amino acid sequence of HSP-1, a major protein isolated from stallion seminal plasma or acid extracts of ejaculated spermatozoa. The protein consists of 121 amino acids organized in two types of homologous repeats arranged in the pattern AA'BB'. Each of the 13-15-residue A-type repeats contains two O-linked oligosaccharide chains. The B-type repeats span 44-47 amino acids each, are not glycosylated, and have the consensus pattern of the gelatin-binding fibronectin type-II module. This domain also occurs in the major bovine seminal plasma heparin-binding proteins PDC-109 (...
