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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.
CD26 and adenosine deaminase interaction: its role in the fusion between horse membrane vesicles and spermatozoa. Membrane vesicles of horse seminal plasma present at their surface a highly specific serine-type protease, dipeptidyl peptidase IV/CD26, a surface antigen known to characterize human prostasomes. Horse sperm cells expressed at their surface A(1) adenosine receptors (A(1)AR) and ecto-adenosine deaminase (ecto-ADA), both detected by immunoblot analysis, whereas CD26 was visualized at the equatorial segment by immunofluorescence microscopy. In addition to CD26, horse membrane vesicles showed ecto-ADA. The fusion process between horse sperm cells and vesicles was evidenced by confocal microscopy, ...
Effect of parturition on levels of vitamins A and E and of beta-carotene in plasma and milk of mares. The objective of this study was to investigate in mares the effect of parturition on plasma and milk levels of retinol, beta-carotene, alpha-tocopherol and cholesterol over 12 weeks around parturition. In blood plasma of horses around parturition an increase of all these components was observed. This increase was most impressive in beta-carotene (P<0.05) and less pronounced for vitamin E, vitamin A and cholesterol. The magnitude of increase around parturition corresponded well with the magnitude of accumulation in colostrum; levels of beta-carotene in colostrum were 65 times higher compared...
New approaches to detect cortisol administration in the horse. The cortisol threshold concentration of 1.0 microg/ml in horse urine adopted by the International Federation of the racing Authorities in 1994 is specific. However, an increase in the sensitivity for the detection of cortisol administration would be helpful. Previous studies have shown that 20beta-dihydrocortisol concentration in urine would be a good indicator of cortisol administration. The purpose of the present work was to estimate the population parameters and the critical values of 20beta-dihydrocortisol and 20beta-dihydrocortisone concentration in urine compared with that of cortisol. U...
PGFM response to exogenous oxytocin and determination of the half-life of oxytocin in nonpregnant mares. We investigated the half-life of oxytocin in reproductively normal mares and the prostaglandin response after oxytocin administrations. Mares were given oxytocin, 10 or 25 iu, i.v., on the day of, or 2 days after, ovulation, and frequent jugular blood samples were collected for analysis of oxytocin and Prostaglandin F metabolite (PGFM) by RIA. Neither dose of oxytocin nor day of treatment affected the half-life of the exogenous oxytocin, which was determined to be 6.8 min. A significant increase in PGFM was observed within 6 min of oxytocin administration and peak values were observed within 1...
Abnormal regulation of muscle contraction in horses with recurrent exertional rhabdomyolysis. To determine whether abnormal regulation of muscle contraction similar to that associated with malignant hyperthermia (MH) was evident in intact external intercostal muscle cells from Thoroughbreds with recurrent exertional rhabdomyolysis (RER). Methods: 5 adult Thoroughbred horses with RER and 7 clinically normal adult Thoroughbred or mixed-breed horses. Methods: Twitch time course variables and contracture responses to various concentrations of potassium, caffeine, and halothane were measured in small bundles of intact external intercostal muscle cells from clinically normal horses and horse...
Pharmacokinetics of carprofen enantiomers in equine plasma and synovial fluid – a comparison with ketoprofen. Carprofen is a Non Steroidal Anti-Inflammatory Drug (NSAID) which is widely used for the treatment of musculoskeletal disorders in horses. The commercial preparation is a racemic mixture of two enantiomers (R and S carprofen). We used HPLC to measure plasma and synovial fluid R and S carprofen concentrations following a single intravenous (i.v.) dose, and computer modelling to determine the pharmacokinetic parameters of the enantiomers in these two body fluids. A comparison was made with results from an identical experiment using ketoprofen. The plasma elimination half lives of R and S carprof...
Pre- and postjunctional effects of inflammatory mediators in horse airways. In addition to their direct contractile effects, histamine (Hist), serotonin [5-hydroxytryptamine (5-HT)], and leukotriene (LT) D(4), in low concentrations, dramatically augment electrical field stimulation (EFS)-induced smooth muscle contractions in equine airways. To determine the mechanism of their action, we studied, in trachealis strips, the effect of these mediators on both cholinergically induced tension and the release of ACh from cholinergic nerves. All three mediators synergistically augmented the contraction of the trachealis that was due to release of endogenous ACh, i.e., EFS-indu...
Different approaches to the identification of a cortisol isomer compound in horse urine. A threshold concentration for cortisol in equine urine was fixed at 1.0 microgram ml-1 by the racing authorities in 1994. In some circumstances, interlaboratory discrepancies were observed and structural cortisol modification was revealed. In order to elucidate the degradation process and to prevent it, an identification study of the produced compound was carried out. The modified substrate was characterised by the same molecular weight as cortisol and a shorter retention time under the conditions used for the cortisol quantification (M.A. Popot, PhD Thesis, King's College, London, 1996). To i...
Nortestosterone: endogenous in urine of goats, sheep and mares? For a number of species it is known that nortestosterone, either the alpha- or beta-epimer, can be of endogenous origin. For goats and mares similar results have not yet been published. As a follow-up on the experiments with cattle, a large number of urine samples per animal were collected from pregnant goats, sheep and mares. These samples were analysed for the presence of alpha- and beta-nortestosterone and alpha-estradiol using GC-MS. The results show that in the goats and mares studied alpha-nortestosterone is present during pregnancy. In this study no alpha-nortestosterone could be demons...
Investigations on the stereoselective action of isoxsuprine on alpha- and beta-adrenoceptors in equine common digital artery. The affinity and functional effects of isoxsuprine enantiomers were investigated to determine the enantiospecificity of the beta-agonistic and alpha-blocking effects. Functional assays on isolated smooth muscle preparations from equine common digital artery were performed to determine the apparent affinity (pD(2)) and intrinsic activity (alpha(E)) of (-)erythro-isoxsuprine (alphaS, betaR, gammaR) and (+)erythro-isoxsuprine (alphaR, betaS, gammaS). The affinity of two enantiomers for the different adrenoceptor types was studied by radioligand binding assays on membrane preparations from the sam...
Effects of anabolic steroid (19-nortestosterone) on the secretion of testicular hormones in the stallion. The aim of this study was to clarify the effect of anabolic steroids on the testicular endocrine function of mature stallions. Mature thoroughbred stallions were treated with 800 mg nandrolone decanoate every 3 weeks for 3 months. After the first treatment, plasma concentrations of LH, immunoreactive inhibin and testosterone decreased rapidly to the nadir. These hormones were maintained at significantly lower concentrations compared with concentrations in intact stallions. Histology of the testicular tissue indicated the arrest of advanced spermatogenesis in the seminiferous tubules and a seve...
Effect of exercise training, selenium and vitamin E on some free radical scavengers in horses (Equus caballus). Physical exercise increases both tissue needs for oxygen and cellular respiration and causes an overproduction of free radicals. When free radical generation exceeds the cell's antioxidant capacity tissue-damage develops due to oxidative stress. Therefore, it appears important to increase the scavenger ability of the tissues. Controlled training and dietary supplements may provide ways of doing this. As a model, we used 3-year-old racehorses (Equus caballus) which underwent a series of different physical exercise trials before and after 70 days of daily training and dietary supplements (vitami...
[HPLC separation and characterization of tick-borne encephalitis and equine Venezuela encephalomyelitis viral proteins]. Homogeneous (according to PAGE) capsid and surface viral proteins were isolated from concentrated purified suspensions of tick-borne encephalitis and Venezuelan equine encephalomyelitis viruses by one-stage reversed-phase HPLC. The amino acid composition and the sequences of their N-terminal parts were determined.
Cannulation in situ of equine umbilicus. Identification by gas chromatography-mass spectrometry (GC-MS) of differences in steroid content between arterial and venous supplies to and from the placental surface. Equine umbilicus was cannulated in utero and a series of cord plasma samples removed for analysis. After steroid extraction and derivatisation, gas chromatographic-mass spectrometric (GC-MS) analysis demonstrated large differences in steroid content between the plasma samples obtained from the umbilical artery and vein, the blood supplies leading to and from the placental surface, respectively. 3Beta-hydroxy-5,7-androstadien-17-one, dehydroepiandrosterone, pregnenolone, 3beta-hydroxy-5alpha-pregnan-20-one, 5-pregnene-3beta,20beta-diol and 5beta-pregnane-3beta,20beta-diol were identified as maj...
The erythrocyte cytoskeleton protein 4.2 is not demonstrable in several mammalian species. Erythrocyte membrane proteins from 44 representative mammals were studied. Protein 4.2 was not detected in guinea pigs (Cavia porcellus) (N = 14), Southern Brazilian swamp large rats (Myocastor coypus) (N = 2), cutias (Dasyprocta sp) (N = 4), and horses (Equus caballus) (N = 13). These animals also presented high ankyrin concentrations except for the horse which did not exhibit a sharp band, although minor components located between proteins 2 and 3 could account for the ankyrin family. The rodents studied did present band 6, which was not detectable in other common rodents such as white rats ...
Analysis of myosin heavy chains at the protein level in horse skeletal muscle. Combined methodologies of enzyme-linked immunosorbent assay (ELISA), sodium dodecyl sulphate polyacrilamide gel electrophoresis (SDS-PAGE), immunoblotting, traditional myofibrillar ATPase (mATPase) histochemistry and immunocytochemistry of whole biopsied samples were used to study myosin heavy chain (MHC) isoforms in the equine gluteus medius muscle. The ELISA technique allowed the quantification of the three MHC isoforms known to be present in different horse muscles: slow (MHC-I) and two fast (termed MHC-IIA and MCH-IIX). The SDS-PAGE method resolved MHCs in three bands: MHC-I, MHC-IIX and M...
A plasma membrane-associated hyaluronidase is localized to the posterior acrosomal region of stallion sperm and is associated with spermatozoal function. Sperm hyaluronidase has been implicated in sperm penetration of the extracellular matrix of the cumulus oophorus and may play a crucial role in gamete interaction and fertility in mammals. The objectives of this study were to characterize the enzyme activity of equine sperm hyaluronidase and to investigate its cellular distribution. Zymography of stallion sperm plasma membrane extracts was used to identify hyaluronidase activity in protein bands. Affinity-purified polyclonal IgG raised against equine sperm hyaluronidase was used to label fresh and capacitated stallion sperm, followed by indire...
Near infrared spectroscopy in large animals: optical pathlength and influence of hair covering and epidermal pigmentation. The effects of epidermal pigmentation and hair covering on the relative transparency of various animal tissues to near infrared (NIR) light were examined, and the pathlengths of NIR light through tissues at four wavelengths in the NIR range were subsequently determined. Black hair covering and black or dark-coloured hooves prevented NIR light from penetration sufficient for conduction of pathlength or NIR spectroscopy measurements. Non-pigmented hair covering of the head did not appear to be a barrier to successful NIR light transmission. Tissues sufficiently transparent to NIR light had the d...
Effects of fetuin on zona pellucida hardening and fertilizability of equine oocytes matured in vitro. In vitro fertilization (IVF) has had poor success in the horse, a situation related to low rates of sperm penetration through the zona pellucida (ZP). Zona pellucida hardening (ZPH) is seen in mouse and rat oocytes cultured in serum-free medium. The hardened ZP is refractory to sperm penetration. Fetuin, a component of fetal calf serum, inhibits ZPH and allows normal fertilization rates in oocytes cultured in the absence of serum. We evaluated whether fetuin is present in horse serum and follicular fluid (FF) and whether fetuin could inhibit ZPH in equine oocytes matured in vitro, thus increas...
A comparison of the bioequivalence of 0.5% fenbendazole top dress pellets or 10% fenbendazole oral suspension against a spectrum of equine parasites. A controlled test was conducted to assess the efficacy bioequivalence of a single dose of 0.5% fenbendazole (FBZ) top dress pellets to a 10% FBZ suspension formulation (Panacur suspension 10%, Hoechst Roussel Vet). Thirty horses with naturally-acquired parasite infections, in replicates of three, were used. Strongyle egg per gram counts were not significantly different (P>0.1) between groups pretreatment, but FBZ treated groups were significantly different from the control group post-treatment. At necropsy, which occurred seven to nine days post-treatment, two methods of nematode recovery were...
[The effect of the essential fatty acids in mare’s milk on the function of the immune system and of nonspecific resistance in rats]. The influence of essential fatty acids of mare's milk fat on the immunocompetent system and nonspecific resistance of male rats weighing 135-145 g was investigated after 6 weeks of feeding. Rats were fed with isocaloric purified diets containing 15% of test fat (in control--combination of lard and sunflower oil) which provide the ratio of omega-6/omega-3 fatty acids equal to 0.76. An increase of immune responsiveness and nonspecific resistance in the group fed with the diet with mare's milk fat on day 6 after a single immunization of the animals with 5% sheep erythrocyte suspension was noted.
Distribution of fast myosin heavy chain-based muscle fibres in the gluteus medius of untrained horses: mismatch between antigenic and ATPase determinants. The distribution of muscle fibres classified on the basis of their content of different myosin heavy chain (MHC) isoforms was analysed in muscle biopsies from the gluteus medius of adult untrained horses by correlating immunohistochemistry with specific anti-MHC monoclonal antibodies and standard myofibrillar ATPase (mATPase) histochemistry. Percutaneous needle biopsies were taken at 3 depths (20, 40 and 60 mm) from 4 4-y-old Andalusian stallions. The percentage of 'pure' I MHC fibres increased whereas that for pure IIX MHC fibres decreased from the most superficial to the deepest sampling sit...
Nitric oxide synthase activity in healthy and interleukin 1beta-exposed equine synovial membrane. To quantitate nitric oxide synthase (NOS) activity in healthy and interleukin 1beta (IL-1beta)-exposed equine synovial membrane. Methods: 6 healthy horses, 2 to 8 years old. Methods: Recombinant human IL-1beta (0.35 ng/kg of body weight) was injected intra-articularly into 1 metacarpophalangeal joint of each horse. The contralateral joint served as an unexposed control. All horses were euthanatized 6 hours after injection of IL-1beta, and synovial membrane specimens were assayed for NOS activity by measuring conversion of arginine to citrulline. Severity of inflammation was semiquantitated by ...
Cimetidine inhibits nitric oxide associated nitrate production in a soft-tissue inflammation model in the horse. Cimetidine (CIM) is an H2-receptor antagonist that has been used in racehorses in an attempt to reduce the occurrence of stress-related gastric ulceration. It has also been shown to produce several useful effects other than its gastric acid suppression properties. Further, it is a well documented antagonist of cytochrome P-450 (CYP) mediated oxygenation reactions. Nitric oxide (NO), a recently discovered mediator or modifier of numerous physiological functions, is generated by several forms of nitric oxide synthase (NOS), one of which is inducible (iNOS). Inducible NOS, expressed in neutrophil...
Independent nucleation and heterogeneous assembly of structure during folding of equine lysozyme. The refolding of equine lysozyme from guanidinium chloride has been studied using hydrogen exchange pulse labelling in conjunction with NMR spectroscopy and stopped flow optical methods. The stopped flow optical experiments indicate that extensive hydrophobic collapse occurs rapidly after the initiation of refolding. Pulse labelling experiments monitoring nearly 50 sites within the protein have enabled the subsequent formation of native-like structure to be followed in considerable detail. They reveal that an intermediate having persistent structure within three of the four helices of the alph...
Etodolac in equine urine and serum: determination by high-performance liquid chromatography with ultraviolet detection, confirmation, and metabolite identification by atmospheric pressure ionization mass spectrometry. A high-performance liquid chromatographic method was used for the detection of etodolac in equine serum and urine. The method consisted of a one-step liquid-liquid extraction, separation on a reversed-phase (RP-18) column and detection using an ultraviolet detector. Additional confirmation methods included a HPLC coupled with an atmospheric pressure chemical ionization mass spectrometer (APCI-MS). Free (unbound) etodolac and its conjugates were present in the samples. Concentrations of the drug in the serum and urine samples collected from four standardbred mares after a single oral administra...
Three-dimensional structure of mare diferric lactoferrin at 2.6 A resolution. Lactoferrin is a monomeric glycoprotein with a molecular mass of approximately 80 kDa. The three-dimensional structure of mare diferric lactoferrin (mlf) has been determined at 2.6 A resolution. The protein crystallizes in the space group P 212121with a=85.2 A, b=99.5 A, c=103.1 A with a solvent content of 55 % (v/v). The structure was solved by the molecular replacement method using human diferric lactoferrin as the model. The structure has been refined using XPLOR to a final R -factor of 0.194 for all data in the 15-2.6 A resolution range. The amino acid sequence of mlf was determined using ...
Oxytocin-neurophysin I mRNA abundance in equine uterine endometrium. A positive-feedback loop between luteal oxytocin and uterine prostaglandin F2 alpha (PGF) is a major signal for luteolysis in ruminants. Likewise, uterine PGF causes luteolysis in mares, but the involvement of oxytocin in this process is unclear. We wanted: 1) to determine if the oxytocin-neurophysin I (OT-NP I) gene is transcribed into mRNA in the endometrium of mares; and, if so, 2) to analyze relative changes in abundance of endometrial OT-NP I mRNA throughout the estrous cycle and during early stages of pregnancy. Endometrial biopsies were obtained from nonbred mares during estrus, and 5, ...
