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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.
Serum osteocalcin and CTX-MMP concentration in young exercising thoroughbred racehorses. Bone responds to exercise with changes in bone (re-)modelling, which might be monitored non-invasively with biochemical bone markers. The aim of this study was to evaluate the influence of exercise on serum osteocalcin and serum carboxy-terminal cross-linked telopeptide of type I collagen generated by matrix metalloproteinases (CTX-MMP) concentration in young racehorses. Seventy-one 2 to 4-year-old Thoroughbreds were included in this prospective infield study. Blood sampling was performed six times (i.e. six sampling cycles) during a 9-month period. Serum samples were analysed with commercial ...
Optimization of assay conditions for leukotriene B4 synthesis by neutrophils or platelets isolated from peripheral blood of monogastric animals. Neutrophils are involved in inflammation through leukotriene (LT) production. The predominant proinflammatory leukotriene released from neutrophils is LTB4, which serves as a biological marker of inflammation. The purpose of this study was to optimize the conditions ex vivo for LTB4 production by neutrophils from horses and dogs, and platelets from chickens. Optimal production of LTB4 was characterized by incubation time (2.5, 5, 10, 15 or 20 min), temperature (25 or 37 degrees C), and calcium ionophore A23187 concentration (0.1, 1, 10 or 20 microM). Incubation longer than 2.5 min did not incr...
Expression of aromatase and oestrogen receptors in reproductive tissues of the stallion and a single cryptorchid visualised by means of immunohistochemistry. Androgen metabolism may proceed to amplify the action of testosterone by its aromatisation to oestradiol. Recently, a growing body of evidence suggests a role of oestrogens in the male reproductive tract via their specific oestrogen receptors (ERs). In order to check whether androgens are converted to oestrogens in the testis, epididymis and prostate of the stallion, the expression of aromatase was visualised by means of immunohistochemistry. Moreover, to show the cellular targets for oestrogens the presence of oestrogen receptor alpha (ERalpha) and oestrogen receptor beta (ERbeta) was demonst...
Dopamine receptors in equine ovarian tissues. Dopamine (DA) agonist and antagonist treatments can affect ovarian reproductive events in the mare. To support our theory that DA produces these effects by acting directly on the ovary, we analyzed equine ovarian tissues for the presence of dopamine receptor-1 (D1r) and dopamine receptor-2 (D2r) mRNA by reverse transcription polymerase chain reaction (RT-PCR) and D1r and D2r proteins by Western blot and immunohistochemistry (IHC). RT-PCR was performed on RNA isolated from ovarian cortex, medulla, granulosa/theca or corpus luteum (CL) tissues and from pituitary (D2r control) and renal artery (D...
Evaluation of developmental changes in the coexpression of myosin heavy chains and metabolic properties of equine skeletal muscle fibers. To determine the growth-related changes in metabolic and anatomic properties in equine muscle fiber type, including hybrid fibers identified with immunohistochemical analysis. Methods: 24 2-, 6-, 12-, and 24-month-old female Thoroughbreds. Methods: Samples were obtained from the gluteus medius muscle of all horses. Expression of myosin heavy chain (MHC) isoforms MHC-I, -IIa, -IIb, and -IIx in each muscle fiber was detected by use of 4 primary monoclonal antibodies: BA-D5, SC-71, BF-F3, and BF-35, respectively. Five muscle fiber types (types I, I/IIA, IIA, IIA/IIX, and IIX) were immunohistochem...
Acute insulin-induced hypoglycaemia does not alter IGF-1 and LH release in cyclic mares. Lactation in the mare is associated with changes in the release of metabolic as well as reproductive hormones. Plasma glucose concentration is constantly reduced in lactating compared with non-lactating mares. Several metabolic signals have been proposed to link nutrition and somatic metabolism with reproductive function. The following experiment was performed to study the effect of acute hypoglycaemia on the release of insulin-like growth factor-1 (IGF-1) and luteinizing hormone (LH) in cyclic mares. Different doses of insulin (0.1 and 0.2 IU/kg body weight) were given to induce a decrease in...
Follicular fluid concentrations of free insulin-like growth factor (IGF)-I during follicular development in mares. The objective of the present study was to evaluate changes in concentrations of free insulin-like growth factor (IGF)-I in follicular fluid (FFL) during follicle development in the mare. Mares (n = 14) were classified as either in the follicular phase (n = 8) or luteal phase (n = 6). Follicles (n = 92) were categorized as small (6-15 mm; n = 54), medium (16-25 mm; n = 23) or large (>25 mm; n = 15) and FFL was collected. Free IGF-I levels in FFL in large follicles of follicular phase mares were greater (P < 0.05) than in large follicles of luteal phase mares and small or medium follicles ...
Nitration and increased alpha-synuclein expression associated with dopaminergic neurodegeneration in equine pituitary pars intermedia dysfunction. Equine pituitary pars intermedia dysfunction (PPID) is a spontaneously occurring progressive disease affecting aged horses and ponies. The pathogenesis of PPID is poorly understood, but the available evidence supports a loss of dopaminergic inhibition of the melanotropes of the pars intermedia. Horses with PPID have increased plasma concentrations of pars intermedia pro-opiomelanocortin-derived peptides that decrease in response to dopamine or dopamine agonist administration. Dopamine and dopamine metabolite concentrations are decreased in the pars intermedia of affected horses compared to age...
Quantitative analysis of voltage-gated potassium currents from primary equine (Equus caballus) and elephant (Loxodonta africana) articular chondrocytes. In this comparative study, we have established in vitro models of equine and elephant articular chondrocytes, examined their basic morphology, and characterized the biophysical properties of their primary voltage-gated potassium channel (Kv) currents. Using whole cell patch-clamp electrophysiological recording from first-expansion and first-passage cells, we measured a maximum Kv conductance of 0.15 +/- 0.04 pS/pF (n = 10) in equine chondrocytes, whereas that in elephant chondrocytes was significantly larger (0.8 +/- 0.4 pS/pF, n = 4, P </= 0.05). Steady-state activation parameters of eleph...
Horse cytokine/IgG fusion proteins–mammalian expression of biologically active cytokines and a system to verify antibody specificity to equine cytokines. Recombinant cytokines are valuable tools for functional studies and candidates for vaccine additives or therapeutic use in various diseases. They can also be used to generate specific antibodies to analyze the roles of different cytokines during immune responses. We generated a mammalian expression system for recombinant cytokines using the equine IgG1 heavy chain constant region as a tag for detection and purification of the expressed cytokine, demonstrated here using equine interferon-gamma (IFN-gamma), interleukin-2 (IL-2), interleukin-4 (IL4) and transforming growth factor-beta1 (TGF-beta1...
Development of an electrospray ionization mass spectrometric method for the quantification of theophylline in horse serum. A rapid and selective method has been developed for the determination of theophylline in horse serum by LC-ESI/MS/MS. The analytical method includes a protein precipitation extraction for sample preparation, liquid chromatography separation technique and ionspray tandem mass spectrometry. The drug was extracted from serum using a protein precipitation with acetonitrile and the supernatants were analyzed using an LC-ESI/MS/MS instrument. The chromatography was performed using a 50 x 2.1 mm C(8) analytical column and an isocratic mobile phase composes of 60:40 acetonitrile-0.5% formic acid in wa...
Amino acid concentrations in blood serum of horses performing long lasting low-intensity exercise. The aim of this work was to evaluate the changes in the concentrations, after two rides different for distance covered, of different amino acids in endurance horses. Blood samples have been collected from horses just before the start, at the top of a steep slope (819 m difference in height) and just at the end of a 32-km endurance ride. A second group, competing in a 72 km endurance ride, has also been sampled immediately before and after the race. In serum samples, the concentrations of alanine, arginine, asparagine, glycine, isoleucine, histidine, leucine, lysine, methionine, ornithine, phen...
Metabolic responses to oral tryptophan supplementation before exercise in horses. This study was conducted to evaluate the effects of oral tryptophan (Trp) supplementation on exercise capacity and metabolic responses in horses. Three horses had to perform an exercise test: a 15-min warm-up followed by a 60-min walk (1.7 m/s, W1), a 10-min trot (3.1 m/s, T1), a second 60-min walk (1.7 m/s, W2), a second 10-min trot (3.1 m/s, T2) and a final 30-min walk (1.7 m/s, W3) until the horses were unwilling to continue. The horses exercised on a treadmill at a 6% incline and with a constant draught load of 40 kg (0.44 kN). Two hours before exercise horses were given 50 g Trp (9.8-10.7...
Effects of draught load exercise and training on calcium homeostasis in horses. This study was conducted to investigate the effects of draught load exercise on calcium (Ca) homeostasis in young horses. Five 2-year-old untrained Standardbred horses were studied in a 4-month training programme. All exercise workouts were performed on a treadmill at a 6% incline and with a constant draught load of 40 kg (0.44 kN). The training programme started with a standardized exercise test (SET 1; six incremental steps of 5 min duration each, first step 1.38 m/s, stepwise increase by 0.56 m/s). A training programme was then initiated which consisted of low-speed exercise sessions (LSE; ...
Influence of the coat color on the trace elemental status measured by particle-induced X-ray emission in horse hair. The influence of hair color on the trace elemental status in horse's hair has been studied. A current analytical technique such as particle-induced X-ray emission (PIXE) used in this study has provided reliable, rapid, easy, and relatively inexpensive diagnostic methods. Twenty-eight elements (Al, Br, Ca, Cl, Co, Cu, Cr, Fe, Ga, Hg, K, Mg, Mn, Mo, Na, Nb, Ni, P, Pb, Rb, S, Se, Si, Sr, Ti, V, Y, and Zn) in mane hair were detected by the PIXE method. The gray hair contains significantly greater amounts of Cu, Ti, and Zn, and lower amounts of Br, Ca, Se, and Sr than those in other colored horse h...
Digital cushions in horses comprise coarse connective tissue, myxoid tissue, and cartilage but only little unilocular fat tissue. Digital cushions were studied in horses with particular reference to vascularization, tissue constituents and matrix components. The cushions mainly resembled a network of coarse collagen bundles. The areas inbetween the bundles were replenished with loosely woven interstitial connective tissue, myxoid tissue, and fibrocartilage. Expected masses of fat lobules were missing: only solitary adipocytes or small groups of adipocytes were seen. Vascular supply to the cushions was remarkably poor. The mucinous myxoid matrix largely consisted of hyaluronan with little sulphated glycosaminoglycans. Myx...
Sperm motility patterns and metabolism in Catalonian donkey semen. The Sperm-Class Analyzer detected four subpopulations of spermatozoa with different motility characteristics in the ejaculate of the Catalonian donkey. Significant differences (P < 0.001) in the distribution of these subpopulations, as well as in total sperm number and percentage total motility, were seen in the diluted semen of four sampled donkeys. All the ejaculates evaluated showed excellent semen quality characteristics; the sperm they contained was more rapid than horse sperm. Principal components analysis showed sperm l-lactate production to be a good predictor of semen condition. Th...
Relationship of seminal plasma level and extender type to sperm motility and DNA integrity. The relationship between seminal plasma level (0, 10, or 20%) and extender type [Kenney type (EZ-Mixin-CST) or Kenney-modified Tyrodes-KMT] to the susceptibility of sperm DNA to denaturation and sperm motility measures were investigated in cooled (5 degrees C) stallion sperm. Three ejaculates from each of three fertile stallions were collected in an artificial vagina and processed as follows: diluted one part uncentrifuged semen with four parts of extender to a final concentration of 20% seminal plasma in either CST or KMT (20% CST; 20% KMT); diluted to a final concentration of 25 million sper...
Characterization of equine intestinal fatty acid binding protein and its use in managing horses with colic. To determine the nucleotide sequence of the equine intestinal fatty acid binding protein (I-FABP) gene, its expression in various regions of the gastrointestinal tract, and the use of measuring I-FABP in horses with colic. Animals-86 horses with colic. Methods: The mRNA sequence for the I-FABP gene was obtained by use of a rapid amplification of complementary DNA ends technique. Comparative I-FABP gene expression was quantitated by use of a real-time reverse transcription-polymerase chain reaction assay. Amounts of I-FABP in abdominal fluid and plasma were measured by use of an ELISA kit. Asso...
Cloning and expression of type III collagen in normal and injured tendons of horses. To clone the 5' end of type III collagen and describe its pattern of mRNA and protein expression in normal and healing tendons in horses. Methods: 14 healthy adult horses. Methods: The tensile region of collagenase-injured superficial digital flexor tendons was harvested at intervals from 1 to 24 weeks after injury. Total RNA was reverse-transcribed into cDNA for cloning and sequencing of type III collagen. Equine-specific nucleic acid probes were developed and used for northern blot analysis and in situ hybridization. Type III collagen protein and cyanogen bromide-cleaved collagen peptides we...
Evaluation of glucose metabolism in three horses with lower motor neuron degeneration. To determine whether increased glucose metabolism is the potential cause of the decreased plasma glucose curve determined after oral glucose tolerance testing in horses with lower motor neuron degeneration. Methods: 3 horses with signs suggestive of lower motor neuron degeneration, 1 horse with malignant melanoma with multiple metastases, and an obese but otherwise healthy horse. Procedures-Glucose metabolism was assessed by use of the hyperglycemic clamp and euglycemic hyperinsulinemic clamp techniques. Results: Mean rate of glucose metabolism of horses with lower motor neuron degeneration wa...
On the difference in stability between horse and sperm whale myoglobins. The work in the literature on apomyoglobin is almost equally divided between horse and sperm whale myoglobins. The two proteins share high homology, show similar folding behavior, and it is often assumed that all folding phenomena found with one protein will also be found with the other. We report data at equilibrium showing that horse myoglobin was 2.1 kcal/mol less stable than sperm whale myoglobin at pH 5.0, and aggregated at high concentrations as measured by gel filtration and analytical ultracentrifugation experiments. The higher stability of sperm whale myoglobin was identified for both...
GPX5 orthologs of the mouse epididymis-restricted and sperm-bound selenium-independent glutathione peroxidase are not expressed with the same quantitative and spatial characteristics in large domestic animals. We report here on the cloning of cDNAs coding bovine and equine orthologs of mouse epididymis-restricted and sperm-bound glutathione peroxidase 5 (GPX5), a selenium-independent member of the multigenic GPX family in mammals. The complete sequence of bovine GPX5 as well as a partial sequence of the equine GPX5 were characterized, conceptually translated and aligned with other known mammalian GPX5 proteins. Using Northern blotting assays, we show that the level of expression of GPX5 is high in bovine but low in equine and that in both species the regionalization of GPX5 expression in epididymis ...
Neuronal nitric oxide synthase is heterogeneously distributed in equine myofibers and highly expressed in endurance trained horses. Mammalian skeletal muscle expresses splice variants of neuronal nitric oxide synthase (nNOS). Skeletal muscles have a metabolically heterogeneous population of myofibers, and fiber composition in equine skeletal muscle is correlated with athletic ability in endurance events. In this study, we investigated whether nNOS expression in equine skeletal muscle is related to fiber type and endurance training. Biopsy samples obtained from the gluteus medius of sedentary- (SH) and endurance-trained (TH) horses were examined for the electrophoretic mobility of myosin heavy chain (MHC) and NOS activity. ...
Binding of cellular proteins to the leader RNA of equine arteritis virus. The genome of equine arteritis virus (EAV) produces a 3' coterminal-nested set of six subgenomic (sg) viral RNAs during virus replication cycle, and each set possesses a common leader sequence of 206 nucleotides (nt) in length derived from the 5' end of the viral genome. Given the presence of the leader region within both genomic and sg mRNAs, it is likely to contain cis-acting signals that may interact with cellular or viral proteins for RNA synthesis. Gel mobility shift assays indicated that proteins in Vero cell cytoplasmic extracts formed complexes with the positive (+) and negative (-) st...
Expression of endothelial and inducible nitric oxide synthases is modulated in the endometrium of cyclic and early pregnant mares. The expression of the endothelial and inducible nitric oxide synthases (eNOS and iNOS, respectively) was examined in the endometrium of cyclic and pregnant mares by real-time polymerase chain reaction and immunohistology. The concentration of eNOS mRNA varied throughout the oestrous cycle, with significantly higher transcripts on Day 5 of the oestrous cycle (P 0.05). In early pregnant mares both eNOS and iNOS mRNA increased between Days 12 and 15 (P < 0.05). In cyclic mares, eNOS protein was detected immunocytochemically in endometrial epithelia, the basement membrane, the endothelial laye...
Muscle metabolic changes associated with long-term inhalation anaesthesia in the horse analysed by muscle biopsy and microdialysis techniques. During anaesthesia in the horse, muscle blood flow has been found to be reduced, possibly leading to hypoxia or ischaemia in the muscle. The aim of this study was to use the muscle biopsy and microdialysis techniques to determine whether long-term inhalation anaesthesia in laterally recumbent horses induces metabolic changes in gluteal muscle indicative of anaerobic metabolism. Muscle biopsies and plasma samples were taken from seven horses at the start and end of halothane anaesthesia. In six isoflurane-anaesthetised horses, given three pharmacological provocations (dobutamine, detomidine, ac...
Differential regulation of the GLUT1 and GLUT3 glucose transporters by growth factors and pro-inflammatory cytokines in equine articular chondrocytes. Glucose serves as the major energy substrate for articular chondrocytes and as the main precursor for the synthesis of extracellular matrix glycosaminoglycans in cartilage. Chondrocytes have been shown to express several glucose transporter (GLUT) isoforms including GLUT1 and GLUT3. The aim of this investigation was to determine the effects of endocrine and cytokine factors on the capacity of equine articular chondrocytes for transporting 2-deoxy-d-[2,6-3H] glucose and on the expression levels of GLUT1 and GLUT3. Chondrocytes maintained in monolayer culture were stimulated for 24 h with TNF-al...
Blood-oxygen binding in healthy Standardbred horses. The purpose of this study was to determine the effect of regulating factors on the oxygen equilibrium curve (OEC) under standard conditions and then to calculate the oxygen extraction between arterial and jugular venous blood in healthy Standardbred horses. The results were compared to those previously obtained in humans and cattle, using the same experimental method. The partial oxygen pressure at 50% saturation of haemoglobin, measured under standard conditions (standard P50), was 24.8+/-2.0 (SD of mean) mmHg. This value was similar to the cattle standard P50 (25.0+/-1.4 mmHg, SD of mean) bu...
