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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.
Kinetics and haematological effects of erythropoietin in horses. A plasma kinetic study of erythropoietin (EPO) was carried out in 4 horses after subcutaneous administration (30 IU/kg bwt) of recombinant human erythropoietin (rhEPO). At standardized intervals for 48 h before injection and for 60 h post-administration, the EPO plasma levels were determined with an immunoradiometric assay based on a sandwich technique. The peak plasma concentration (30-113 mIU/ml) was observed after a delay ranging from 6 to 9 h post-administration and the drug levels reached a physiological value around 60 h following rhEPO injection. Moreover, reference values for plasma EP...
Equine pituitary neoplasia: a clinical report of 21 cases (1990-1992). The records of 21 horses with an adenoma of the pars intermedia of the pituitary gland were reviewed. The animals comprised nine Dutch warmblood horses and 12 ponies. They ranged in age from 12 to 30 years (mean +/- sem 21 +/- 1.3 years) for 20 of the horses, and comprised 13 geldings and eight mares. All the animals showed hirsutism. In 1991, nine horses were diagnosed as having the adenoma, equivalent to 0.5 per cent of the horses examined in 1991. The mean +/- sem survival time of four of the horses (from discharge to the development of complications which required euthanasia) was 192 +/- 5...
Equine plasminogen polymorphism: allelic frequencies in 23 breeds. A modified procedure for detection of the two alleles of equine plasminogen using Western blotting methods following polyacrylamide gel isoelectric focusing is described. Gene frequencies in 23 breeds and Equus przewalskii are provided.
Comparative study of the oxyhaemoglobin dissociation curve of four mammals: man, dog, horse and cattle. 1. The entire oxygen dissociation curve (ODC) and the effects of temperature, pH and 2,3-diphosphoglycerate (DPG) on this curve, have been compared in four mammalians: man, dog, horse and cattle. 2. If the oxyphoric capacities are similar between these species (around 1.39 ml O2/gHb), their P50, measured in standard conditions, i.e. at pH 7.4; pCO2 40 mmHg and T 37 degrees C, varies between 23.8 (+/- 0.8) mmHg for the horse, 25.0 (+/- 1.4) mmHg for cattle, 26.6 (+/- 1.2) for man and 28.8 (+/- 2.6) mmHg for the dog. 3. The higher dispersion of the dog's P 50 is due to difference between breeds;...
Kinetic analysis of D-xylose absorption after its intragastric administration to mares deprived of food. Multicompartmental analysis was applied to study the kinetics of D-xylose distribution after its intragastric administration to healthy mares deprived of food for 12, 36, 72, and 96 hours. Disposition of D-xylose was described by a 5-compartment model. Maximal plasma D-xylose concentration was similar for 12 and 36 hours of food deprivation and was greater (P = 0.0001) than the values for 72 and 96 hours. Peak concentration of D-xylose appeared progressively later as food deprivation proceeded (P = 0.0001). Fractional rate of transfer (k1,6) was less after 96 hours of food deprivation, compare...
A comparison of the reduction in immunoglobulin (IgG) concentration of frozen equine plasma treated by three thawing techniques. The IgG concentration of plasma from 13 mares was measured by radial immunodiffusion when fresh and after storage at -4 degrees C and thawing by 3 methods. There was no significant (P > 0.05) reduction in the IgG concentration when plasma was thawed over 6 hours at 22 degrees C (1352.9 +/- 101.6 mg/dL) (mean +/- SEM) compared with the fresh sample (1369.5 +/- 88.1 mg/dL). In contrast, there was a significant (P < 0.05) reduction in the IgG concentration of plasma that was rapidly thawed at 57 degrees C over 50 minutes (1142.9 +/- 66.2 mg/dL), or placed in a microwave oven for 20 to 25 minutes ...
Aromatase activity in the mare ovary during estrous cycle. Measurement of endogenous steroids and of their in vitro inhibitory effect. This present study was undertaken to clarify estrogen synthesis in the mare ovary. First of all, an evaluation of endogenous steroid contents was carried out in the follicular fluid and in the luteal tissue at different stages of the luteal phase. Radioimmunoassays were performed after separation and purification of each hormone by chromatography. High amounts of conjugated (0.9 mg/l) and unconjugated (4 mg/l) estradiol-17 beta were found in the follicular fluid of the large follicules (50 mm). These concentrations of estrogens decreased drastically in the luteal tissue, and only low levels of...
Determination of alclofenac in equine plasma and urine by high-performance liquid chromatography. A high-performance liquid chromatographic method to measure plasma and urinary alclofenac levels in equine biofluids is described. Isolation of the drug from plasma is achieved using liquid-liquid extraction with diethyl ether. Reversed-phase C18 solid phase extraction is used for the extraction of free and conjugated alclofenac from urine. The reproducibility and accuracy of the method were well within acceptable limits over the concentration ranges 0-10 and 0-20 micrograms/ml, respectively, for plasma and urine. Starting with 2 ml of plasma, a concentration of 0.1 microgram/ml could easily b...
Partially folded states of equine lysozyme. Structural characterization and significance for protein folding. Despite their homologous structure, c-type lysozymes and alpha-lactalbumins have been found to differ profoundly in their unfolding behavior, in that the alpha-lactalbumins readily enter a partially unfolded collapsed state (the "molten globule"), whereas lysozymes unfold cooperatively to a highly unfolded state. The calcium-binding property of lysozyme from equine milk provides an evolutionary link between the two families of proteins. We demonstrate here that equine lysozyme undergoes a two-stage unfolding transition upon heating or in the presence of guanidine hydrochloride that is highly d...
Yield and composition of milk from lactating mares: effect of body condition at foaling. The yield and composition of milk from nursing mares were studied during the first 2 months of lactation in two groups of six and five saddle mares respectively made thin or fat before foaling, then fed ad lib. post partum. Milk yield did not significantly vary with body condition (16.5 and 15.4 kg/d for fat and thin mares respectively). Milk from fat mares had a higher fat content than milk from thin mares; the reverse was found for protein. The composition of milk fat was modified: milk of fat mares was poorer in short- and medium-chain fatty acids (C8-C14). Milk fat output was affected by b...
Biochemical and haematological changes following prolonged halothane anaesthesia in horses. Six healthy horses were anaesthetised with halothane (1.2 times the horse minimal alveolar concentration) in oxygen for more than 12 hours. Serum bilirubin, aspartate aminotransferase, alkaline phosphatase and L-iditol dehydrogenase values were significantly (P < 0.05) increased for up to nine days after anaesthesia. These changes suggest an anaesthesia related liver dysfunction. Creatine kinase increased to an average of more than 1400 IU litre-1 24 hours after anaesthesia and this change is indicative of muscle cell disruption. Renal-associated biochemical results, (that is serum creatinine ...
Pulsatile secretion of luteinizing hormone and progesterone in mares during the estrous cycle and early pregnancy. Existence of ultradian variation in serum progesterone concentration and the relation between progesterone and luteinizing hormone (LH) secretory patterns were investigated in nonpregnant and pregnant mares. Blood samples were taken every 15 minutes for a 24-hour period on day 8 of the estrous cycle and day 18 of pregnancy, respectively. Progesterone and LH concentrations were determined by radioimmunoassay. Progesterone was secreted in pulsatile manner in nonpregnant and pregnant mares. Luteinizing hormone also was secreted in a pulsatile manner in both groups of mares. There was little tempo...
Accumulation of allantoin and uric acid in plasma of exercising trotters. Plasma concentrations of hypoxanthine, uric acid, and allantoin, which are breakdown products of adenine nucleotides, were measured in Standardbred and Finnhorse trotters during and after an exercise test on a high-speed treadmill, after an incremental exercise test performed on a racetrack, and after a racing competition. Fiber-type composition of the middle gluteal muscle and the muscle concentrations of adenine nucleotides and inosine monophosphate were measured after the racetrack test. Changes in the concentration of hypoxanthine were not observed in any of the tests. Peak concentration o...
Different resistance of mammalian red blood cells to hemolysis by bile salts. To evaluate why hemolysis of red blood cells (RBC) by bile acids varies in different mammalian species, we determined the mean corpuscular volume (MCV), lipid content and the concentrations of the conjugates of deoxycholate and of NaCl inducing 50% hemolysis of RBC from healthy humans, pigs, horses, cows, sheep and jaundiced humans. A volume of 0.05 mL of washed RBC at 1% hematocrit, which has the same lipid content but different phospholipid composition and number of erythrocytes (owing to the variable MCV), was incubated in taurodeoxycholate (TDC) solution (0-5 mM) to determine the TDC conce...
Developmentally regulated changes in the glycoproteins of the equine embryonic capsule. The embryonic capsule, which covers the equine blastocyst after it loses its zona pellucida, is composed of mucin-like glycoproteins. In the present study, we investigated both macroscopic and molecular changes in the capsule during development. The weight of the capsule increased from day 11-12 of pregnancy and reached a maximum at about day 18, coinciding with the time during which the conceptus migrates extensively throughout the uterus. The sialic acid content of the capsule declined markedly from about day 16, the time of conceptus 'fixation' in the uterus, which suggests a unique develop...
Effect of heparin on capacitation/acrosome reaction of equine sperm. The onset of sperm capacitation/acrosome reaction was evaluated using heparin. Equine semen was incubated at 38 degrees C for 4.5 h in culture medium with and without 10 micrograms/mL heparin and with and without 0.1 microM of Ca2+ ionophore. Sperm acrosome reaction was detected using chlortetracycline fluorescence (CTC) and transmission electron microscopy (TEM). The CTC assay provided staining patterns that corresponded with the capacitation/acrosome reaction in other mammalian species (man, mouse, guinea pig). The percentages of incapacitated sperm (PUC), capacitated acrosome-intact sperm (...
Comparison of immunoaffinity chromatography combined with gas chromatography-negative ion chemical ionisation mass spectrometry and radioimmunoassay for screening dexamethasone in equine urine. A comparison of the sensitive analytical methods of radioimmunoassay (RIA) and immunoaffinity chromatography (IAC) combined with gas chromatography-negative ion chemical ionisation mass spectrometry for the specific and reliable screening of dexamethasone in equine post-race urine is presented. Results from analyses of samples collected from a mare during 144 hours post administration of 26 mg of dexamethasone sodium phosphate are described.
Rapid high-performance liquid chromatographic method for the determination of ketamine and its metabolite dehydronorketamine in equine serum. A simple, rapid and sensitive high-performance liquid chromatographic procedure has been developed for the determination of ketamine and dehydronorketamine in equine serum. Sample preparation consisted of mixing equal volumes of serum and acetonitrile-phosphoric acid (85%)-water (20:2:78, v/v/v), followed by ultrafiltration through a 10,000 molecular mass cut-off filter. Separation of these two analytes in the ultrafiltrate was accomplished on a reversed-phase phenyl column eluted with methanol-acetonitrile-phosphate buffer solution. Ketamine and dehydronorketamine were detected by a variable ...
Expression of an evolutionarily conserved function associated molecule on sheep, horse and cattle natural killer cells. Natural killer (NK) cells are large granular lymphocytes that lyse a wide variety of transformed and virally-infected target cells without prior exposure to antigen, and without restriction by major histocompatibility complex antigens. Although NK cells have been identified in a variety of mammalian species, how NK cells recognize antigen and trigger lysis is unknown. Recently, monoclonal antibodies made against NK-like cells from teleost fish were shown to react with NK cells from humans and rats, and to inhibit their cytolytic activity. The role of this apparently evolutionarily conserved fu...
Characterization of equine oviductal proteins synthesized and released at estrus and at day 4 after ovulation in bred and nonbred mares. Proteins synthesized and released in vitro by oviducts collected from horse mares during estrus and at day 4 after ovulation for bred and nonbred mares were examined by two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (2-D SDS PAGE) and fluorography. Ampullary and isthmic regions both produced a wide array of nondialyzable proteins in culture. Major proteins or groups of proteins identified according to relative molecular weight (kDa) and apparent isoelectric point (pI) were at 100 kDa, pI 8; 100-200 kDa, pI 6; 150 kDa, pI 4.5; 60-100 kDa, pI 4; and an array of polypep...
Comparison of IgE-binding antigens in horse dander and a mixture of horse hair and skin scrapings. Extracts of horse dander (HD) and horse hair and skin scrapings (HHSS) have been compared with respect to their content of proteins and carbohydrates. The protein content of HD is more than double that of HHSS, while the carbohydrate content is of the same order. SDS-PAGE and IEF, both combined with immunoblotting, and CIE/CRIE showed the IgE-binding ability of the proteins/glycoproteins present in the two extracts. SDS-PAGE/immunoblotting showed the presence of mainly the same IgE-binding bands in the two extracts. Nine were detected in HD, and seven in HHSS. Four of these were glycoproteins....
Investigation of stimulus-secretion coupling in equine sweat gland epithelia using cell culture techniques. When sweat glands isolated from samples of horse skin were explanted and cultured under favourable conditions, they could exhibit cellular outgrowth. This growth could be maintained for 2-4 weeks and these primary cultures were then disaggregated and the resultant cell suspensions used to initiate epithelial cell lines. Secretion from intact equine sweat glands is regulated by beta 2-adrenoceptors and appears to be mediated by cyclic AMP, but there is evidence that calcium may also play a role. Adrenaline could increase the cyclic AMP content of the cultured cells and this response was mediate...
Plasma lipid transport in the horse (Equus caballus). 1. Equine plasma contains lipoproteins corresponding to very low density (VLDL), low density (LDL) and high density lipoproteins (HDL). 2. HDL accounts for approximately 60% of plasma lipoprotein mass and consists of a single population of particles. 3. LDL is heterogeneous comprising three discrete subfractions. 4. Two proteins are found in the region of apolipoprotein (apo) B-100 in VLDL and LDL and a third similar to apoB-48 is in VLDL. 5. Lecithin:cholesterol acyl transferase is active in plasma and hepatic lipase and lipoprotein lipase are evident in post-heparin plasma. 6. There is no si...
The disposition of suxibuzone in the horse. A high performance liquid chromatographic method is described to determine the anti-inflammatory drug suxibuzone (SXB) and its major metabolites phenylbutazone (PBZ) and oxyphenbutazone (OPBZ) in equine plasma and urine. When suxibuzone (6 mg/kg) was administered intravenously (i.v.) or orally (p.o.) no parent drug was detected in plasma or in urine. The disposition of the metabolite PBZ (i.v.) could be described by a 2 compartment model with a beta half-life varying from 7.40 to 8.35 h. Due to severe side effects the use of i.v. suxibuzone should not be encouraged in the horse. PBZ and OPBZ w...
Detection of bicarbonate administration (milkshake) in standardbred horses. Total plasma carbon dioxide (TCO2) concentrations were measured in Standardbred horses to determine criteria to discriminate between normal horses and horses with excessive TCO2 concentrations on raceday. TCO2 concentrations from stabled horses were distributed normally with a mean of 30.2 mmol/L and a standard deviation of 1.2 (n = 192) while pre-race TCO2 concentrations were not normally distributed. The results indicate that about 50 horses per million are likely to have TCO2 concentrations greater than or equal to 35 mmol/L and that it is extremely unlikely that a normal horse would have a...
Purification and characterization of equine complement factor C3. A rapid method for purifying equine C3 which yields milligram quantities of pure C3 is described. Protein from equine plasma was selectively precipitated with polyethylene glycol, and the C3 was purified by anionic and cationic exchange HPLC. The yield from this procedure was 12%. The purified C3 was composed of an alpha chain (118 kD) and a beta chain (68 kD) linked by at least one disulfide bond, and it had an isoelectric point of 4.7. Amino acid analysis indicated a strong conservation of amino acid usage between equine and human C3. The N-terminal sequences of the alpha and beta chains wer...
Furosemide magnifies the exercise-induced elevation of plasma vasopressin concentration in horses. The purpose of this study was to test the hypothesis that furosemide administration before exercise would cause greater increases in plasma arginine vasopressin (AVP) concentration in exercising horses than exercise alone. Six adult, clinically normal, unfit mares underwent three randomly ordered 60 minute standard exercise tests on an equine treadmill to examine the effect of furosemide administration on plasma AVP concentration. In one trial, furosemide (1 mg kg-1) was infused four hours before exercise (FUR-4) and a placebo (10 ml saline) was infused two minutes before exercise; in another ...
Independent modulation of horse airway smooth muscle by epithelium and prostanoids. The effects of epithelial removal and cyclooxygenase inhibition on contractions induced by exogenous acetylcholine (ACh) and electrical field stimulation (EFS) were evaluated in horse tracheal strips and bronchial rings. Epithelial removal potentiated the response to ACh but had no influence on the response to EFS. The effect of epithelial removal was not altered by pretreating the tissues with meclofenamate, a cyclooxygenase inhibitor. In trachealis strips, meclofenamate augmented contractions induced by EFS but not by ACh. In bronchial rings, meclofenamate augmented EFS-induced contraction t...
Twenty-four hour gastric pH monitoring and blood gastrin concentrations in fasted ponies. Gastric pH varied widely among eight fasted ponies, of which seven exhibited discrete episodes of spontaneous alkalinisation (SA). SA occurred at all times of the study and no significant variation in the summary variables of pH (median pH, mean pH and percentage of readings exceeding pH 4.0) was noted among the periods 0 to eight, eight to 16 and 16 to 24 hours. The occurrence of SA has significant implications for the performance of acid secretory studies in the fasted pony. There was, however, no significant correlation between pH and plasma gastrin concentration measured using a commercial...
