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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.
Stereospecific pharmacokinetics of free and protein-bound ketoprofen in serum and synovial fluid of horses after intravenous and intramuscular administration. To determine intravascular and intrasynovial pharmacokinetics of the R and S enantiomers of ketoprofen after i.v. and i.m. administration to horses. Methods: 6 healthy adult mares. Methods: Horses were weighed and ketoprofen (2.2 mg/kg of body weight) was administered i.v. Blood and synovial fluid samples were obtained and analyzed for concentrations of the R and S enantiomers by means of a modified reverse-phase stereospecific high-pressure liquid chromatographic method. Three weeks later, the procedure was repeated, except that ketoprofen was given IM. Protein binding of ketoprofen enantiome...
Intrafollicular insulin-like growth factor-binding protein levels in equine ovarian follicles during preovulatory maturation and regression. The profiles of insulin-like growth factor-binding proteins (IGFBPs) in follicular fluid have been characterized in a number of mammals (rats, pigs, sheep, cattle, humans) and are good indicators of follicular status. We studied the IGFBP profiles of equine serum and ovarian follicular fluid recovered at various stages of the follicular phase. The levels of IGFBPs were related to the morphology and the steroidogenic activity of the follicles. Follicular fluids were recovered by ultrasound-guided follicular aspiration. In the first experiment, the dominant follicles of 10 mares were partly punc...
Effect of copper supplementation on copper status of pregnant mares and foals. Pregnant Thoroughbred mares (n = 21) were grazed on tall fescue pasture containing 4.4-8.6 mg Cu/kg dry matter (DM). Twelve mares were supplemented with approximately 0.5 mg Cu/kg liveweight (LW)/day as copper sulphate for the final 13-25 weeks gestation. Plasma copper concentration declined during the third trimester for all mares and was not affected by copper supplementation. Liver copper concentration of mares and foals was determined when the foals were 4-10-days-old. Copper supplementation of the mares tended to increase mare liver copper concentration (P<0.08) and increased foal live...
Effect of copper supplementation on the copper status of pasture-fed young Thoroughbreds. The effect of copper supplementation of pasture fed mares and foals on the copper status of the foals, in terms of plasma, soft tissue and bone copper concentrations and caeruloplasmin activity, was investigated. Twenty-one Thoroughbred foals from either control mares (n = 9), or copper-supplemented mares (n = 12) were divided randomly into control (pasture only, n = 10) or supplemented (pasture and oral copper sulphate, n = 11) groups. The pasture diet was grazed by all animals, and contained 4.4-8.6 mg Cu/kg dry matter (DM). The copper supplement for the mares contained copper sulphate equiv...
[Lysozyme activity in the milk of sucking mares during lactation]. It was the aim of this project to investigate the changes of the lysozyme activity in the milk of mares during the lactation period. Further on the influence of race, date of conception and foaling, age and number of lactations on the lysozyme activities in milk was analysed. Milk samples were collected from 44 mares (trotters, warmblood, quarter horses) from eight farms between the 1st and 90th day p. p. The activity of the lysozyme was measured by a turbidometric method. Summarizing the following results are obtained: Lysozyme activities in mare milk of the 1st and 3rd day p. p. were higher ...
Role of cAMP and neuronal K+ channels on alpha 2-AR-induced inhibition of ACh release in equine trachea. To investigate the effects of changes in intracellular cAMP on alpha 2-adrenoceptor (AR)-induced inhibition of airway acetylcholine (ACh) release, we examined the effects of the alpha 2-AR agonist clonidine on electrical field stimulation-evoked ACh release from equine tracheal parasympathetic nerves before and after treatment with 8-bromo-cAMP or forskolin. We also tested whether charybdotoxin (ChTX)- or iberiotoxin (IBTX)-sensitive Ca(2+)-activated K+ channels mediate alpha 2-AR-induced inhibition by examining the effect of clonidine in the absence and presence of ChTX or IBTX on ACh release...
Stereochemical structures of synthesized and natural plasmalogalactosylceramides from equine brain. Modified galactosylceramide with a long-chain cyclic acetal at the sugar moiety, plasmalogalactosylceramide, was isolated from equine brain. To identify the isomeric stereostructure of the natural product, the plasmalo derivative was chemically synthesized from galactosylceramide through acetalization. The presence of cyclic acetal linkage, the linked position and length of the acetal chain of the synthesized and natural products were determined by proton nuclear magnetic resonance spectroscopy and fast-atom bombardment-mass spectrometry, as well as gas chromatography-mass spectrometry and gas...
Characterization of the haemolytic activity of Streptococcus equi. The haemolytic activity of Streptococcus equi, the cause of equine strangles, was characterized. Production of haemolysin in Todd Hewitt broth was dependent on an equine serum supplement and the logarithmic phase of growth after which activity declined sharply. RNA core also induced haemolysin production from cells harvested at the end of the logarithmic phase of growth. Haemolysis was not affected by cholesterol, was only slightly increased in reducing conditions and was completely inactivated by trypan blue, identifying the haemolytic activity as streptolysin S-like (SLS-like). Purification ...
A conserved structural element in horse and mouse IGF2 genes binds a methylation sensitive factor. The equine IGF2 gene has been cloned and characterised. It spans a 9 kb region, which is substantially less than the corresponding human gene. Three coding exons and three untranslated leader exons, all highly homologous to those in other species, were identified. Downstream of the polyadenylation site in exon 6, a dinucleotide repeat sequence was identified. Three putative promoters (P1-P3) were localised in the 5' region of the gene. RNase protection analysis revealed two active promoters in fetal tissues, P2 and P3, whereas P3 was the only promoter active in adult tissues. This represents a...
Equine infectious anemia virus transactivator is a homeodomain-type protein. Lentiviral transactivator (Tat) proteins are essential for viral replication. Tat proteins of human immunodeficiency virus type 1 and bovine immunodeficiency virus form complexes with their respective RNA targets (Tat responsive element, TAR), and specific binding of the equine anemia virus (EIAV) Tat protein to a target TAR RNA is suggested by mutational analysis of the TAR RNA. Structural data on equine infectious anemia virus Tat protein reveal a helix-loop-helix-turn-helix limit structure very similar to homeobox domains that are known to bind specifically to DNA. Here we report results of...
Further characterization of equine brain gangliosides: the presence of GM3 having N-glycolyl neuraminic acid in the central nervous system. Equine brain gangliosides were isolated and their structures were characterized, to examine whether equine brain has N-glycolyl neuraminic acid in gangliosides, since other mammals predominantly possess N-acetyl neuraminic acid in brain gangliosides, and equine erythrocytes and organs except the brain have gangliosides exclusively containing N-glycolyl neuraminic acid. The gangliosides purified from the brain were identified by proton NMR spectroscopy and mass spectrometry, as well as GLC, resulting in their identification as GM4, GM3, GM2, GM1, GD1a, GD1b, and GT1b. Of these gangliosides, GM3...
Contribution of K+ channels and ouabain-sensitive mechanisms to the endothelium-dependent relaxations of horse penile small arteries. 1. Penile small arteries (effective internal lumen diameter of 300 600 microm) were isolated from the horse corpus cavernosum and mounted in microvascular myographs in order to investigate the mechanisms underlying the endothelium-dependent relaxations to acetylcholine (ACh) and bradykinin (BK). 2. In arteries preconstricted with the thromboxane analogue U46619 (3-30 nM), ACh and BK elicited concentration-dependent relaxations, pD2 and maximal responses being 7.71+/-0.09 and 91+/-1 % (n=23), and 8.80+/-0.07 and 89+/-2% (n=24) for ACh and BK, respectively. These relaxations were abolished by me...
The effects of equivalent doses of tromethamine or sodium bicarbonate in healthy horses. To describe the effects of tromethamine, a putative treatment for metabolic acidosis, and to compare its biochemical effects with those of sodium bicarbonate. Methods: Randomized intervention study with repeated measures. Methods: 16 healthy horses, 3 to 17 years old, weighing 391 to 684 kg. Methods: Ten horses received 3 mEq/kg tromethamine and six received 3 mEq/kg sodium bicarbonate. Samples of venous blood and cerebrospinal fluid (CSF) were collected at intervals before and after drug administration. Heart rate and breathing rate were also recorded at intervals. Results: Median standard ba...
Dietary control of exertional rhabdomyolysis in horses. To determine whether feeding a low-carbohydrate, high-fat diet would decrease severity of exercise-induced muscle injury in horses with exertional rhabdomyolysis. Methods: 19 horses with a history of exertional rhabdomyolysis. Methods: Case series. Methods: Specimens of the semitendinosus or semimembranosus muscle were obtained for histologic examination, and serum creatine kinase (CK) and aspartate transaminase (AST) activities 4 hours after exercise were determined. Horses were then fed a low-carbohydrate, high-fat diet, and serum CK and AST activities 4 hours after exercise were reevaluated...
Peculiarities of vitamin D and of the calcium and phosphate homeostatic system in horses. The aim of the present study was to investigate the importance of putative regulatory factors of the calcium (Ca) and inorganic phosphate (P(i)) homeostatic system in the horse. The concentrations of Ca, P(i), vitamin D metabolites, parathyroid hormone (PTH), the activity of the alkaline phosphatase (AP) and the concentration and binding properties of vitamin D binding protein (DBP) were measured in the plasma. In addition, the ability of the renal cortex to hydroxylate calcidiol into 24,25(OH)2D3 and 1,25(OH)2D3 was evaluated in vitro. The plasma concentration of Ca (3.2 +/- 0.15 mmol.L-1, N ...
HPLC on Chiralcel OJ-R for enantiomer separation and analysis of ketoprofen, from horse plasma, as the 9-aminophenanthrene derivative. Racemic ketoprofen is a non-steroidal anti-inflammatory drug used to treat musculoskeletal and colic conditions in horses. The enantioselective chiral inversion of ketoprofen administered to horses has been studied by use of cellulose tris(4-methylbenzoate), also known as Chiralcel OJ-R, as chiral stationary phase; acetonitrile - 0.02 M perchlorate buffer (pH 2.0)-methanol, 60:15:25 (v/v/v) was used as mobile phase. Before chromatography, to effect adequate chiral interaction with the chiral stationary phase ketoprofen was derivatized with 9-aminophenanthrene, under acid conditions, after soli...
The effect of propanediol on the morphology of fresh and frozen equine embryos. Seventeen horse embryos recovered on the sixth day after spontaneous ovulation were; 1) washed in PBS (n = 6), 2) treated with 1.5 M 1-2 propanediol (n = 6) or, 3) frozen and thawed using 1.5 M propanediol as the cryoprotectant (n = 5). After treatment, the embryos were incubated for 6 h in medium before they were fixed, serially sectioned and examined microscopically to count the total numbers of interphase, mitotic and pycnotic nuclei. Significant differences were measured only in the mean proportions of pycnotic cells (+/- s.d.), both between the control (9.2 +/- 7.3%) and frozen-thawed emb...
A comparison of the biochemical composition of equine follicular fluid and serum at four different stages of the follicular cycle. Samples of blood and follicular fluid were recovered from 27 Welsh Pony mares at 4 distinct stages of follicular development. Eighteen biochemical parameters were measured in each sample, including sodium, potassium, chloride, glucose, urea, creatinine, calcium, inorganic phosphate, total bilirubin, total protein, albumin, magnesium, triglyceride, total cholesterol, nonesterified fatty acids, alkaline phosphatase, gamma-glutamyltransferase and aspartate aminotransferase. The concentrations of progesterone, 17beta oestradiol and testosterone, pH and osmolarity, were also measured in all the fol...
Effect of anti-freeze protein (AFP) on the cooling and freezing of equine embryos as measured by DAPI-staining. Equine embryos recovered on Day 6 after ovulation were cooled to +4 degrees C, or frozen with AFP alone or together with glycerol. Twenty embryos (140-200 microm in diameter) were randomly assigned to 6 treatment groups. In the first 3 groups, the embryos were cooled from room temperature to +4 degrees C at a rate of 3 degrees C/min and warmed again at a rate of 32 degrees C/min in a programmable freezer. In the second 3 groups, the embryos were frozen using a standard protocol, stored in liquid nitrogen for 5-7 days and then thawed in a 37 degrees C waterbath. After cooling/warming or freezin...
Lectin-staining pattern in extratesticular rete testis and ductuli efferentes of prepubertal and adult horses. This study was undertaken to determine the lectin affinity of the extratesticular rete testis and ductuli efferentes epithelial cells in adult and prepubertal horses, using ten different lectin horseradish peroxidase conjugates: Con-A, LCA, WGA, GSA-II, SBA, PNA, RCA-I, DBA, UEA-I, and LTA. In some cases, treatments with sialidase and KOH preceded the lectin staining. In sexually mature and immature horses the results showed the presence of different kinds of sialoglycoconjugates with the terminal sialic acid linked to D-GalNAc and beta-D-Gal residues in the rete testis. In the apical surface ...
Activation of extracellular matrix metalloproteinases in equine laminitis. Samples of connective tissue obtained from the hoof of six laminitic and eight non-laminitic adult horses were analysed zymographically to investigate whether connective tissue matrix metalloproteinases are activated or induced during laminitis. The activity or matrix metalloproteinases was substantially greater in the tissues from the laminitic horses than in the tissues from the non-laminitic horses. A comparison of the collagenolytic activity in the laminitic and control tissues showed that collagenolytic activities corresponding to the 92 kDa (P < 0.001), 72 kDa (P < 0.01) and 66 kDa...
[Activities of the enzymes LDH, gamma-GT, GOT, GPT and lactoperoxidase in the milk of breeding mares during the course of lactation]. Milk samples were collected from 44 mares (trotters, warm blood horses, quarter horses) during lactation between the 1st and 90th day p.p. at 20 defined days. The activity of the enzymes LDH, gamma-GT, GOT, GPT and lactoperoxidase was investigated. The aim of this study was to find out the changes of these parameters during lactation and whether an influence of race, conception, date of foaling, age and number of lactations existed on the enzyme activities in mare's milk. The following results were obtained: In mare's milk the LDH-activity was highest (xg = 629 x 1.5 +/- 1 U/l) on the 1st day ...
Development and use of an enzyme-linked immunosorbent assay to monitor serum and urine acepromazine concentrations in thoroghbreds, and possible changes associated with exercise. To develop an ELISA that is sensitive and suitable for measurement of immunoreactive acepromazine (ACP) in horse serum and urine and to determine the acute effects of exercise on immunoreactive ACP values in Thoroughbreds. Methods: 12 healthy Thoroughbreds (5 mares, 5 geldings, 2 stallions), aged 2 to 8 years. Methods: A commercially available antibody and a horseradish peroxidase-conjugated oxime derivative of immunoreactive ACP were used to develop a one-step ELISA. Horses were used in a crossover design study to evaluate possible effects of treadmill exercise on serum and urine ACP concentr...
Chromogenic assays for equine coagulation factors VII, VIII:C, IX, and X, and C1-esterase inhibitor. To adapt manual human chromogenic assays for coagulation factors VII (F.VII), VIII:coagulant (F.VIII:C), IX (F.IX), and X (F.X), and C1-esterase inhibitor (C1-INH) for use with an automated analyzer, and to measure the activity of these proteins in horses. Methods: 10 healthy horses were used to determine ranges for the assays. Pooled plasma for standards was collected from an additional 20 healthy horses. Methods: A computer-assisted analyzer was programmed from the manual method for commercially available human F.VII, F.VIII:C, F.IX, F.X, and C1-INH chromogenic assay kits. Standards were pre...
Interindividual variation in total and carrier-mediated lactate influx into red blood cells. To study in standardbred horses interindividual variation in the influx of lactate into red blood cells, venous blood samples were collected from 89 horses from 2 wk to 9 yr of age. For 62 horses, the rate of influx was normally distributed with a mean rate of 4.09 nmol.mg protein-1.min-1 at a lactate concentration of 10 mM, and the respective value for the other 27 horses was 0.58 nmol.mg protein-1.min-1. At 30 mM of lactate, the rates were 8.71 and 1.97 nmol.mg protein-1.min-1, respectively. This bimodal distribution was independent of age. In horses with high transport activity, the monocar...
Adenosine and hypoxanthine transport in horse erythrocytes: evidence for a polymorphism in the transport of hypoxanthine via a sodium-dependent cotransporter. The inward transport of two purines, adenosine and hypoxanthine, at 37 degrees C by horse erythrocytes was compared. No mediated transport of adenosine was detected in horse erythrocytes, nor was saturable, high-affinity binding of the potent facilitated-diffusion inhibitor nitrobenzylthioinosine demonstrable in horse erythrocyte membranes. In contrast, erythrocytes from most horses possessed a saturable sodium-dependent hypoxanthine transporter (apparent K(m), 100 +/- 28 microM; Vmax, 0.20 +/- 0.08 mmol (l cells)-1 h-1; means +/- S.E.M., n = 5). Guanine inhibited hypoxanthine influx (apparent...
Catecholamine and cortisol responses of horses to incremental exertion. The responses of the plasma concentrations of catecholamines and cortisol in horses to varied relative intensities of exertion were examined. The plasma concentrations of cortisol, epinephrine and norepinephrine increased significantly (p < 0.05) with exertion. The plasma cortisol concentrations at relative work intensities of 48.3% +/- 1.4%, 82.3% +/- 2.0% and 99.6% +/- 0.4% of VO2max were 114%, 124%, and 126%, respectively, of those at rest, whereas the plasma epinephrine concentrations were 239%, 772% and 3483%, and the norepinephrine concentrations were 138%, 255%, and 1121% of the valu...
Physiology of body fluids in the horse. This article presents a brief overview of the physiology of homeostasis in the horse and discusses the units of measurement by which dissolved electrolytes are assessed. Differences in electrolyte composition between compartments of the body and the manner in which water is exchanged between them are reviewed. The mechanisms by which alterations in osmolality and effective circulating volume are detected and the physiological responses to those alterations are presented.
Effect of exercise on acid-base status of horses. Exercise in horses is associated with a wide variety of physiological changes in fluid, electrolyte and acid base balance. The integration of physiologic and physiochemical mechanisms acts to minimize alterations in pH and enhance removal of carbon dioxide produced by exercising muscles. This article provides a description of the changes that take place during exercise and how these changes affect acid-base balance in the horse.
