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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.
Quantification of 19-nortestosterone sulphate and boldenone sulphate in urine from male horses using liquid chromatography/tandem mass spectrometry. Following administration of the anabolic steroid 19-nortestosterone or its esters to the horse, a major urinary metabolite is 19-nortestosterone-17beta-sulphate. The detection of 19-nortestosterone in urine from untreated animals has led to it being considered a naturally occurring steroid in the male horse. Recently, we have demonstrated that the majority of the 19-nortestosterone found in extracts of 'normal' urine from male horses arises as an artefact through decarboxylation of the 19-carboxylic acid of testosterone. The aim of this investigation was to establish if direct analysis of 19-n...
Effects of methylprednisolone acetate and glucosamine on proteoglycan production by equine chondrocytes in vitro. To evaluate the effects of methylprednisolone acetate (MPA) on proteoglycan production by equine chondrocytes and to investigate whether glucosamine hydrochloride modulates these effects at clinically relevant concentrations. Methods: Articular cartilage with normal gross appearance from metacarpophalangeal and metatarsophalangeal joints of 8 horses (1 to 10 years of age). Methods: In vitro chondrocyte pellets were pretreated with glucosamine (0, 1, 10, and 100 microg/mL) for 48 hours and exposed to MPA (0, 0.05, and 0.5 mg/mL) for 24 hours. Pellets and media were assayed for proteoglycan prod...
Analysis of major proteins and fat fractions associated with mare’s milk fat globules. Although several studies have aimed to identify mare's milk proteins, only the major whey proteins and some caseins have yet been characterized. Incomplete sequencing of the equine genome and the difficulty of recovering highly hydrophobic proteins mean that little is known to date about the proteins associated with milk fat globules, which have been shown to play an important role in newborns' defense mechanisms. The fat fraction, in particular the distribution of unsaturated fatty acids, has been more extensively studied, but complex lipids are only partially elucidated. This study reports a...
Equine placenta expresses glutamine synthetase. In most mammalian species the developing fetus utilizes large amounts of glutamine derived both from the maternal circulation and synthesized de novo in the placenta. The present study was designed to determine the role of the placenta in glutamine synthesis in the horse. The placentae from eight Standardbred mares were sampled immediately after parturition together with additional tissues obtained at necropsy from three Standbred mares during diestrous. Glutamine synthetase protein was detectable in the non-pregnant horn of the placenta in amounts similar to those seen in gluteus muscle, but ...
Quantitative intradermal terbutaline sweat test in horses. The aim of the current study was to quantify sweating responses to intradermal terbutaline in normal horses. Seven Thoroughbred horses were used. Terbutaline (10-fold dilutions from 1000-0.001 mg/l) and a saline control were injected intradermally (0.1 ml/site) and sweat collected for 30 min into absorbent pads taped over each injection site. Tests were performed monthly for 11 successive months and temperature, relative humidity and dewpoint were measured at the time of testing. There was no significant effect (P<or=0.05) of environmental variables or time of year on sweat responses at any...
Development of a liquid chromatography-tandem mass spectrometry method for an euthanasic veterinarian drug: Tanax. A development of a rapid and sensitive LC-MS/MS method for the simultaneous detection of active ingredients of the euthanasic veterinarian drug Tanax mixture is described. The method proposed, with a retention time of few minutes (6 min) was developed for an equine serum sample with solid-phase extraction (S.P.E). This S.P.E. procedure has been revealed useful for the determination of very low concentrations of Tanax analytes (0.05-1 ng/ml). The method was validated in terms of specificity/selectivity, sensitivity, recovery and precision.
Fat content and fatty acids profile of colostrum and milk of primitive Konik horses ( Equus caballus gmelini Ant.) during six months of lactation. The effect of the stage of lactation, the number of foals and age of the mare on changes in the fat content and fatty acid composition of colostrum and milk of primitive Konik horses was investigated. Colostrum and milk samples from 12 lactating mares were collected at the beginning of lactation, on the days 1 and 2 after foaling and then, starting from the first month of lactation, at 4-week intervals up to the sixth month of lactation. Significant differences were observed in fat content as well as the composition of some analysed fatty acids between colostrum and milk of mares of the Konik ...
Actions of a nitric oxide donor on prostaglandin production and angiogenic activity in the equine endometrium. Nitric oxide (NO) plays an important role in prostaglandin secretion and angiogenesis in the reproductive system. In the present study, the roles of the NO donor spermine NONOate and tumour necrosis factor-alpha (TNF; as a positive control) in prostaglandin production and angiogenic activity of equine endometria during the oestrous cycle were evaluated. In addition, the correlation between NO production and the expression of key prostaglandin synthase proteins was determined. The protein expression of prostaglandin F synthase (PGFS) increased in early and mid-luteal stages, whereas that of pro...
Proteins associated with the early intrauterine equine conceptus. A critical period of early gestation in the mare involves the immobilization (fixation) of the encapsulated conceptus at around days 16-17. We compared the major proteins in the normal equine embryonic capsule and endometrial secretions around the period of fixation with those from pregnancies in the process of termination induced by administration of an analogue of prostaglandin F(2 alpha) (PGF(2 alpha)). Uterocalin and beta(2)-microglobulin (beta(2)M) associated with the embryonic capsule were proteolytically converted to smaller forms during the fixation period. These conversions were simil...
Complementary DNA cloning, functional expression and characterization of a novel cytochrome P450, CYP2D50, from equine liver. Members of the CYP2D family constitute only about 2-4% of total hepatic CYP450s, however, they are responsible for the metabolism of 20-25% of commonly prescribed therapeutic compounds. CYP2D enzymes have been identified in a number of different species. However, vast differences in the metabolic activity of these enzymes have been well documented. In the horse, the presence of a member of the CYP2D family has been suggested from studies with equine liver microsomes, however its presence has not been definitively proven. In this study a cDNA encoding a novel CYP2D enzyme (CYP2D50) was cloned f...
Dynamics of pruning waste and spent horse litter co-composting as determined by chemical parameters. Co-composting of pruning waste and horse manure was monitored by different parameters. A windrow composting pile, having the dimensions 2.5m (height) x 30m (length) was established. The maturation of pruning waste and horse manure compost was accompanied by a decline in NH(4)(+)-N concentration, water soluble C and an increase in NO(3)(-)-N content. Organic matter (OM) content during composting followed a first-order kinetic equation. This result was in agreement with the microbiological activity measured by the CO(2) respiration during the process. The correlation at a high level of probabili...
Characterization and quantification of fluoxymesterone metabolite in horse urine by gas chromatography/mass spectrometry. Fluoxymesterone, an anabolic steroid with the 17alpha-methyl,17beta-hydroxy group, has been developed as an oral formulation for therapeutic purposes. However, it is also used illegally in racehorses to enhance racing performance. In this study, we detected 9alpha-fluoro-17,17-dimethyl-18-norandrostane-4,13-dien-11beta-ol-3-one by gas chromatography/mass spectrometry (GC/MS), which has not been reported as a fluoxymesterone metabolite so far in horse. It was synthesized for use as a reference standard, and characterized on the basis of (1)H NMR and (13)C NMR spectra, as well as GC/MS EI mass s...
Intrafollicular Concentrations of Steroid Hormones and PGF2α in Relation to Follicular Development in the Mares during the Breeding Season. The concentrations of androstenedione, estradiol-17β, progesterone and PGF2α contained in the follicular fluid produced by the follicles in collected ovaries of mares that have had estrous phase during the breeding season were measured and analyzed the relation between the growth stage of follicles and the hormone levels in the follicular fluid. An ultrasonographic diagnostic instrument was used to measure the diameter of the follicles in order to categorize the follicles into three groups the following: 8 small follicles (from 1.0 to less than1.5 cm), 8 medium follicles (from 1.5 to less th...
Elucidation of the MD-2/TLR4 interface required for signaling by lipid IVa. LPS signals through a membrane bound-complex of the lipid binding protein MD-2 and the receptor TLR4. In this study we identify discrete regions in both MD-2 and TLR4 that are required for signaling by lipid IVa, an LPS derivative that is an agonist in horse but an antagonist in humans. We show that changes in the electrostatic surface potential of both MD-2 and TLR4 are required in order that lipid IVa can induce signaling. In MD-2, replacing horse residues 57-66 and 82-89 with the equivalent human residues confers a level of constitutive activity on horse MD-2, suggesting that conformational...
Effect of adipose-derived nucleated cell fractions on tendon repair in horses with collagenase-induced tendinitis. To assess the potential of adipose-derived nucleated cell (ADNC) fractions to improve tendon repair in horses with collagenase-induced tendinitis. Methods: 8 horses. Methods: Collagenase was used to induce tendinitis in the superficial digital flexor tendon of 1 forelimb in each horse. Four horses were treated by injection of autogenous ADNC fractions, and 4 control horses were injected with PBS solution. Healing was compared by weekly ultrasonographic evaluation. Horses were euthanatized at 6 weeks. Gross and histologic evaluation of tendon structure, fiber alignment, and collagen typing were...
Hair analysis of anabolic steroids in connection with doping control-results from horse samples. Doping control of anabolic substances is normally carried out with urine samples taken from athletes and horses. Investigation of alternative specimens, e.g. hair samples, is restricted to special cases, but can also be worthwhile, in addition to urine analysis. Moreover, hair material is preferred in cases of limited availability or complicated collection of urine samples, e.g. from horses. In this work, possible ways of interpretation of analytical results in hair samples are discussed and illustrated by practical experiences. The results demonstrate the applicability of hair analysis to det...
High-intensity exercise induces structural, compositional and metabolic changes in cuboidal bones–findings from an equine athlete model. Fatigue fracture of cuboidal bones occurs in the human foot as well as the equine carpus. The racehorse provides a naturally-occurring model to study the effects of high-intensity exercise on the morphology and metabolism of cuboidal bones. We studied both the mineral and the collagenous matrix of the third (C(3)) and radial (C(r)) carpal bones of raced and non-raced Thoroughbred (TB) horses. We hypothesised that racehorses would show increases in the mineral component of these bones and post-translational modifications of the collagenous matrix alongside changes in markers of collagen remodel...
Effect of feeding increasing quantities of starch on glycaemic and insulinaemic responses in healthy horses. The aim of this study was to investigate the effect of increasing the intake of starch on the glycaemic and insulinaemic responses of horses. A cross-over study design was used in which four horses were fed increasing amounts of a compound feed (0.5-3.5 kg) to provide 0.3, 0.6, 0.8, 1.1, 1.4 and 2 starch/kg bodyweight (BW)/meal. The glycaemic response increased with starch intake (P<0.05), while feeding <1.1 g starch/kg BW resulted in a lowered response, compared to when 1.1-2 g starch/ kg BW was fed (P<0.01). The results suggested that insulin responses may be more appropriate to def...
Molecular characterization and expression of the equine M(1) and M(2)-pyruvate kinase gene. To elucidate the molecular properties of the equine glycolytic enzymes equine M(1) (eM(1)) and M(2) (eM(2))-pyruvate kinase (PK), mRNAs were isolated from thoroughbred horse skeletal muscle and hair roots, respectively. The full-length eM(1) and eM(2)-PK cDNAs consist of 2,320 and 2,376 bp, respectively, containing a 1596 bp open reading frame. The cDNAs were mapped to equine chromosome 1, and the equine pyruvate kinase M (PKM) gene consists of twelve exons. Exon 9 of eM(1)-PK and exon 10 of eM(2)-PK were further investigated in five equine species. Out of 55 amino acids encoded by exon 9 in e...
Comparative activities of semicarbazide-sensitive amine oxidase (SSAO) in five domestic species. Semicarbazide-sensitive amine oxidase (SSAO) activity was measured spectrophotometrically using benzylamine as a substrate, in the serum of healthy males and females of horses, camels, cattle, sheep and goats. The animals were born and raised in the same area, and the blood collection was made on the same day to avoid variations. Also the concentrations of protein and copper were measured in the same samples. There were no significant gender-related differences in SSAO activity between the tested animals regardless of species. Activities of SSAO in either male or female of horse were significa...
Immunolocalisation of the uterine secretory proteins uterocalin, uteroferrin and uteroglobin in the mare’s uterus and placenta throughout pregnancy. Previous studies have shown that the equine uterus produces many progesterone-dependent proteins throughout gestation. In particular, uterocalin and uteroferrin are detectable using electrophoresis or blot analyses but information regarding the immunohistochemical placental distribution of these two proteins is rare and information regarding uteroglobin is still lacking. The aim of the present study was to co-immunolocalise these three secretory proteins in the mare's uterus throughout gestation in an effort to understand their functional role in the maintenance of pregnancy. Therefore, endome...
Insulin sensitivity in Belgian horses with polysaccharide storage myopathy. To determine insulin sensitivity, proportions of muscle fiber types, and activities of glycogenolytic and glycolytic enzymes in Belgians with and without polysaccharide storage myopathy (PSSM). Methods: 10 Quarter Horses (QHs) and 103 Belgians in which PSSM status had been determined. Methods: To determine insulin sensitivity, a hyperinsulinemic euglycemic clamp (HEC) technique was used in 5 Belgians with PSSM and 5 Belgians without PSSM. Insulin was infused i.v. at 3 mU/min/kg for 3 hours, and concentrations of blood glucose and plasma insulin were determined throughout. An i.v. infusion of g...
Assessment of vitamin E concentrations in serum and cerebrospinal fluid of horses following oral administration of vitamin E. To determine concentrations of alpha-tocopherol in serum and CSF of healthy horses following administration of supplemental vitamin E in feed. Methods: 10 healthy adult horses. Methods: Horses were allocated to receive supplemental d-alpha-tocopherol (1,000 U/d [group A; n=5] or 10,000 U/d [group B; 5]) in feed for 10 days. Blood samples were collected before (baseline), during, and at intervals for 10 days after discontinuation of vitamin E administration for assessment of serum alpha-tocopherol concentration. Cerebrospinal fluid samples were collected prior to and 24 hours after cessation of...
Analysis of iridoids from Harpagophytum and eleutherosides from Eleutherococcus senticosus in horse urine. LC/ESI-MS n methods have been previously set up to detect the administration of (i) Harpagophytum and (ii) preparations containing a plant capable of anti-stress properties: Eleutherococcus senticosus. Harpagoside has been found to be the main indicator of Harpagophytum administration in the horse. These methods have been applied to a large number of horse urine samples of various origins. Regarding the detection of Harpagophytum administration, harpagoside, harpagide and 8-para-coumaroyl harpagide were detected together in only one sample out of 317. Eleutheroside E was found to be the main i...
Comparative structural and chemical studies of ferritin cores with gradual removal of their iron contents. Transmission Electron Microscopy (TEM), X-ray Absorption Near Edge Spectroscopy (XANES), Electron Energy-Loss Spectroscopy (EELS), Small-Angle X-ray Scattering (SAXS), and SQUID magnetic studies were performed in a batch of horse spleen ferritins from which iron had been gradually removed, yielding samples containing 2200, 1200, 500, and 200 iron atoms. Taken together, findings obtained demonstrate that the ferritin iron core consists of a polyphasic structure (ferrihydrite, magnetite, hematite) and that the proportion of phases is modified by iron removal. Thus, the relative amount of magneti...
Estrogenic activity of the equine estrogen metabolite, 4-methoxyequilenin. Oxidative metabolism of estrogens has been associated with genotoxicity. O-methylation of catechol estrogens is considered as a protective mechanism. 4-Methoxyequilenin (4-MeOEN) is the O-methylated product of 4-hydroxyequilenin (4-OHEN). 4-OHEN, the major catechol metabolite of the equine estrogens present in the most widely prescribed hormone replacement therapeutics, causes DNA damage via quinone formation. In this study, estrogen receptor (ERa) binding of 4-MeOEN was compared with estradiol (E2) and equilenin derivatives including 4-BrEN using computer modeling, estrogen response element (...
Spelt, an ancient cereal and first pressure linseed oil as ingredients of compound feedstuffs for modern horse feeding. Spelt is a covered cereal with large glumellas. In experiment 1, it has been compared in terms of chemical composition with barley, oat and maize. Spelt is characterized by rather low protein and ether extract (EE) contents. The neutral detergent fibre (NDF) content of spelt was slightly higher than that of oat but the acid detergent fibre (ADF) content was lower. Two compound feedstuffs were fed along with hay to six horses used in a cross-over design. Both diets were well appreciated by the horses and there were no significant differences in the apparent digestibility coefficients, except fo...
