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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.
A natural energy absorbent polymer composite: The equine hoof wall. The equine hoof has been considered as an efficient energy absorption layer that protects the skeletal elements from impact when galloping. In the present study, the hierarchical structure of a fresh equine hoof wall and the energy absorption mechanisms are investigated. Tubules are found embedded in the intertubular matrix forming the hoof wall at the microscale. Both tubules and intertubular areas consist of keratin cells, in which keratin crystalline intermediate filaments (IFs) and amorphous keratin fill the cytoskeletons. Cell sizes, shapes and IF fractions are different between tubular a...
Markers for oxidative stress in the synovial fluid of Thoroughbred horses with carpal bone fracture. Arthritis is thought to cause oxidative stress in synovial fluid in humans, but there have been few reports in horses. To evaluate oxidative stress in synovial fluid in horses, this study used 19 horses with unilateral fracture of the carpal joint bone. Synovial fluid was collected from the carpal joint on the fracture (arthritis group) and contralateral (control group) sides. Diacron-reactive oxygen metabolites (d-ROMs) and biological antioxidant potential (BAP) were then measured, and the oxidative stress index (OSI) was calculated. d-ROMs and OSI of the arthritis group were significantly hi...
Elucidation of the biosynthetic pathways of boldenone in the equine testis. Boldenone is an anabolic-androgenic steroid that is prohibited in equine sports. Urine from the uncastrated male horse contains boldenone that is thought to be of endogenous origin and thus a threshold ('cut-off') concentration has been adopted internationally for free and conjugated boldenone to help distinguish cases of doping from its natural production. The testis is likely to be a source of boldenone. Qualitative analysis was performed on extracts of equine testicular homogenates (n = 3 horses) incubated non-spiked and in the presence of its potential precursors using liquid chromatog...
Enantioselective capillary electrophoresis for pharmacokinetic analysis of methadone and 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine in equines anesthetized with ketamine and isoflurane. An enantioselective assay for the determination of methadone and its main metabolite 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine in equine plasma based on capillary electrophoresis with highly sulfated γ-cyclodextrin as chiral selector and electrokinetic analyte injection is described. The assay is based on liquid/liquid extraction of the analytes at alkaline pH from 0.1 mL plasma followed by electrokinetic sample injection of the analytes from the extract across a buffer plug without chiral selector. Separation occurs cationically at normal polarity in a pH 3 phosphate buffer containin...
Pharmacokinetics and selected pharmacodynamics of morphine and its active metabolites in horses after intravenous administration of four doses. The objective of the current study was to describe and characterize the pharmacokinetics and selected pharmacodynamic effects of morphine and its two major metabolites in horses following several doses of morphine. A total of ten horses were administered a single intravenous dose of morphine: 0.05, 0.1, 0.2, or 0.5 mg/kg, or saline control. Blood samples were collected up to 72 hr, analyzed for morphine, and metabolites by LC/MS/MS, and pharmacokinetic parameters were determined. Step count, heart rate and rhythm, gastrointestinal borborygmi, fecal output, packed cell volume, and total prote...
Lipid classes in adipose tissues and liver differ between Shetland ponies and Warmblood horses. Fatty acids, as key components of cellular membranes and complex lipids, may play a central role in endocrine signalling and the function of adipose tissue and liver. Thus, the lipid fatty acid composition may play a role in health status in the equine. This study aimed to investigate the fatty acid composition of different tissues and liver lipid classes by comparing Warmblood horses and Shetland ponies under defined conditions. We hypothesized that ponies show different lipid patterns than horses in adipose tissue, liver and plasma. Six Warmblood horses and six Shetland ponies were housed an...
Potent inhibitors of equine steroid isomerase EcaGST A3-3. Equine glutathione transferase A3-3 (EcaGST A3-3) belongs to the superfamily of detoxication enzymes found in all higher organisms. However, it is also the most efficient steroid double-bond isomerase known in mammals. Equus ferus caballus shares the steroidogenic pathway with Homo sapiens, which makes the horse a suitable animal model for investigations of human steroidogenesis. Inhibition of the enzyme has potential for treatment of steroid-hormone-dependent disorders. Screening of a library of FDA-approved drugs identified 16 out of 1040 compounds, which at 10 μM concentration afforded at ...
Melanocyte-Stimulating Hormone Response to Exercise, Twitching, Epinephrine Injection, Substance P Injection, and Prostaglandin-F2α Administration in Mares. Five experiments were performed to test the hypothesis that α-melanocyte-stimulating hormone (MSH) is secreted in response to various stressors in horses similar to prolactin, growth hormone, and adrenocorticotropin (ACTH). There was considerable variation in resting concentrations of MSH and in the degree of stimulation in responders; thus all data sets were tested for heterogeneity of variance and corrected for as needed before analysis. In experiment 1, 12 mares were used in a switchback design to test the effect of a 2-minute exercise bout on MSH secretion. Plasma MSH concentrations were ...
Expression, localization, and concentration of A-kinase anchor protein 4 (AKAP4) and its precursor (proAKAP4) in equine semen: Promising marker correlated to the total and progressive motility in thawed spermatozoa. A-kinase anchor protein 4 (AKAP4) is playing a central role in flagellar structure, chemotaxis, capacitation and sperm motility. In mammals, AKAP4 is expressed during spermatogenesis. AKAP4 is synthesized as a precursor, proAKAP4, which is cleaved into mature AKAP4 during fibrous sheath assembly. The proAKAP4 is a good indicator of sperm quality in humans and boars. The aims of this work were to study the expression, the localization and the concentration of proAKAP4 and AKAP4 in equine semen, and to evaluate the possible correlation between the total and progressive motility and the concentra...
Cellular and molecular mechanisms of the preovulatory follicle differenciation and ovulation: What do we know in the mare relative to other species. Terminal follicular differentiation and ovulation are essential steps of reproduction. They are induced by the increase in circulating LH, and lead to the expulsion from the ovary of oocytes ready to be fertilized. This review summarizes our current understanding of cellular and molecular pathways that control ovulation using a broad mammalian literature, with a specific focus to the mare, which is unique in some aspects of ovarian function in some cases. Essential steps and key factors are approached. The first part of this review concerns LH, receptors and signaling, addressing the descripti...
The health enhancer yeast Saccharomyces cerevisiae in two types of commercial products for animal nutrition. The health enhancer yeast Saccharomyces cerevisiae (SC) is widely used in diets for different animals. Two main types of SC-based products are commercially available, one containing live yeasts and one containing SC fermentation by-products, which are supposedly not dependent on live yeasts for their physiological effects in vivo. Culture-based techniques were applied to study yeasts in two types of commercial products: a product containing live SC (LSC) and a SC fermentation product (SCFP). Three temperatures (25, 30 and 39°C) and two pH levels (4 and 7) were tested. The product with LSC co...
An optimized TaqMan real-time PCR method for authentication of ASINI CORII COLLA (donkey-hide gelatin). In this study, probe/primers of high specificity and sensitivity were selected to analyze donkey-hide gelatin for donkey DNA and to look for horse, ox, and pig DNA as possible adulterants. The mitochondrial CO I genes in donkey, horse, and ox were selected as target sequences for design and synthesis of three pairs of specific probes and primers. In addition, eight pairs of probe/primers were obtained via literature search. Out of these eleven groups of probe/primers, those with the highest specificity and sensitivity were selected, which was fulfilled by the screening firstly with animal hide...
Equine platelet concentrate preparation and validation. Development of equine platelet concentrate (PC) would aid management of cases requiring transfused platelets (PLTs), where adminstration of whole-blood or platelet-rich plasma (PRP) might be contraindicated. Objective: To test and validate a method for production of an equine PRP-PC product. Methods: Six healthy Thoroughbred geldings from a research herd. Methods: In this prospective experimental study, whole blood was collected and processed through multiple centrifugation steps to yield 120 mL of PC. The PC was stored at 22°C and gently and continuously agitated. Measurements of PLT count...
Analysis of magnetization transfer (MT) influence on quantitative mapping of T2 relaxation time. Multi-echo spin-echo (MESE) protocol is the most effective tool for mapping T relaxation in vivo. Still, MESE extensive use of radiofrequency pulses causes magnetization transfer (MT)-related bias of the water signal, instigated by the presence of macromolecules (MMP). Here, we analyze the effects of MT on MESE signal, alongside their impact on quantitative T measurements. Study used 3 models: in vitro urea phantom, ex vivo horse brain, and in vivo human brain. MT ratio (MTR) was measured between single-SE and MESE protocols under different scan settings including varying echo train lengths, n...
The Effect of Autologous Protein Solution on the Inflammatory Cascade in Stimulated Equine Chondrocytes. Cartilage injury occurs commonly in equine athletes, often precipitating posttraumatic osteoarthritis (PTOA). Orthobiologics such as autologous conditioned serum (ACS) and autologous protein solution (APS) may be useful in decreasing posttraumatic inflammation, thereby preventing PTOA. The objective of this study was to quantify cytokine concentrations in ACS and APS and evaluate the protective effects of ACS and APS on inflamed chondrocytes cultured . We hypothesized that the combination of platelet-derived growth factors (PDGF) and anti-inflammatory cytokines present in APS would be superior...
Seasonal variation in equine follicular fluid proteome. Proteomic studies of follicular fluid (FF) exist for several species, including the horse; however, the seasonal influence on FF proteome has not been explored in livestock. The application of high-throughput proteomics of FF in horse has the potential to identify seasonal variations of proteins involved in follicle and oocyte growth. Methods: This study (i) profiles the proteomes of equine FF collected from dominant growing follicles during the spring anovulatory season (SAN), and spring (SOV), summer (SUM), and fall (FOV) ovulatory seasons; and (ii) identifies season-dependent regulatory net...
Determination of Salivary Cortisol in Donkey Stallions. Salivary cortisol provides information about free plasma cortisol concentration, and salivary sampling is a noninvasive well-tolerated procedure. The aim of this study was to validate a commercial enzyme immunoassay for the determination of salivary cortisol in donkeys. Saliva samples were collected in four donkey stallions on 13 nonconsecutive days at 8:30 AM to avoid circadian variation. Animals were already accustomed to be handled. Saliva was collected using a swab inserted at the angle of the lips, placed onto the tongue for 1 minute and returned into a polypropylene tube. Tubes were cen...
Omega-3 fatty acids and docosahexaenoic acid oxymetabolites modulate the inflammatory response of equine recombinant interleukin1β-stimulated equine synoviocytes. Omega-3 fatty acid (n-3 PUFA) supplementation may have beneficial effects in certain chronic diseases, potentially including osteoarthritis. Favorable effects are attributed, in part, to downstream pro-resolving oxylipid metabolites. We investigated the role of n-3 PUFA and docosahexaenoic acid (DHA)-derived oxylipids (docosanoids) on equine synoviocyte metabolism. We hypothesized that n-3 PUFA and selected docosanoids would modulate inflammatory mediator gene expression by recombinant equine (re)IL-1β-stimulated synovial fibroblasts. Synoviocyte monolayer cultures were prepared from grossly ...
Conformational plasticity of the VEEV macro domain is important for binding of ADP-ribose. Venezuelan equine encephalitis virus (VEEV) is a new world alphavirus which can be involved in several central nervous system disorders such as encephalitis and meningitis. The VEEV genome codes for 4 non-structural proteins (nsP), of which nsP3 contains a Macro domain. Macro domains (MD) can be found as stand-alone proteins or embedded within larger proteins in viruses, bacteria and eukaryotes. Their most common feature is the binding of ADP-ribose (ADPr), while several macro domains act as ribosylation writers, erasers or readers. Alphavirus MD erase ribosylation but their precise contributi...
The efficacy and safety of velagliflozin over 16 weeks as a treatment for insulin dysregulation in ponies. A previous six-week (wk) study demonstrated the potential of the sodium-glucose linked transport inhibitor velagliflozin as a novel treatment for equine insulin dysregulation. The present study examined the safety and efficacy of velagliflozin over 16 wk. of treatment, and over 4 wk. of withdrawal. Twenty-four insulin dysregulated ponies were selected, based on their hyper-responsiveness to a diet challenge meal containing 3.8 g non-structural carbohydrates (NSC)/kg bodyweight (BW). Ponies with serum insulin > 90 μIU/mL either 2 or 4 h after feeding were enrolled, and randomly alloca...
Dexamethasone downregulates expression of several genes encoding orphan nuclear receptors that are important to steroidogenesis in stallion testes. Glucocorticoids impair testosterone synthesis by an unknown mechanism. Stallions treated with the synthetic glucocorticoid dexamethasone had testes collected at 6 or 12 hours postinjection. The testicular expression of selected genes encoding nuclear receptors and steroidogenic enzymes was measured. At 6 hours, dexamethasone treatment decreased levels of NR0B2, NR4A1, NR5A1, and NR5A2 messenger RNAs (mRNAs) and NR5A2 mRNA levels remained depressed at 12 hours. In contrast, dexamethasone increased levels of NFKBIA mRNA at both time points. At 6 hours, dexamethasone did not alter levels ...
Evidence for Increased Content of PGF2α, PGE2, and 6-keto-PGF1α in Endometrial Tissue Cultures From Heavy Draft Mares in Anestrus With Endometritis. Endometritis is one of the most important causes of infertility in mares. Mares may suffer from endometritis outside the breeding season; however, pathological condition is often undiagnosed in anestrus. The aim of this study was to examine whether the secretion profiles of prostaglandin F2α (PGF2α), prostaglandin E2 (PGE2), and a metabolite of prostacyclin I2 (6-keto-PGF1α) differ in endometrial tissue cultures of heavy draft mares in anestrus with endometritis compared to those without endometritis. The endometrial biopsies were collected from 51 heavy draft mares. Inclusion criteria for ...
Influence of mechanical fluid shear stress on the osteogenic differentiation protocols for Equine adipose tissue-derived mesenchymal stem cells. Cell-based therapies have become a promising approach to promote tissue regeneration and the treatment of musculoskeletal disorders. Bone regeneration maintains bone homeostasis, mechanical stability and physical performance. Mechanical stimulation showed to induce stem cell differentiation into the osteogenic fate. However, the effect of various osteogenic protocols on the osteogenic commitment of equine adipose-derived stem cells is not fully elucidated. Here we examined the influence of fluid-based shear stress (FSS) via mechanical rocking to assess whether mechanical stimulation promotes o...
Dynamic Change of Serum Levels of Some Branched-Chain Amino Acids and Tryptophan in Athletic Horses After Different Physical Exercises. Physical exercise induces several adaptations involving modulation of the energetic pathways to meet the energy demand during physical exercise. The branched-chain amino acids, leucine in particular, are considered as the most relevant amino acids, especially for exercise physiology. In this study, the change of leucine, valine, and tryptophan concentration was investigated in athletic horses before and after different exercise type. A total of 40 sport horses were equally divided into 4 groups according to the exercise type: group A (jumping), group B (reining), group C (32 km ride), and grou...
Evaluation of semen characteristics, oxidative stress, and biochemical indices in Arabian horses of different ages during the hot summer season. Egypt is anticipated to be potentially influenced by the global climate warming. Therefore, the current study aimed to evaluate the influence of age on the fertility potential of Arabian stallions during summer breeding months. Arabian horses grouped according to their age into three groups, each involved six stallions: young (5-6 years), middle (11-12 years) and old (15-20 years) age groups, were weekly sampled during the months of July-August. Ejaculates were collected using artificial vagina, Missouri model, and examined for pH, volume, concentration, motility, livability and morphological ...
Detection of ESAs in equine urine and blood by SAR-PAGE. Erythropoiesis-stimulating agents (ESAs) have been used in horses for doping purposes to increase the performance of these animals in endurance sports. Currently, enzyme-linked immunosorbent assay (ELISA) and mass spectrometry methods are used to detect ESA abuse in equines. However, the sarcosyl polyacrylamide gel-electrophoresis (SAR-PAGE) technique could also be used, since its application in human doping control is well established and has proven to be more sensitive. In this work, the SAR-PAGE method was used to detect recombinant human erythropoietin (rHuEPO), novel erythropoiesis stimul...
Pharmacokinetics of meloxicam after oral administration of a granule formulation to healthy horses. Nonsteroidal anti-inflammatory drugs are administered in horses for several systemic diseases. Selective cyclooxygenase-2 inhibitors are preferred because of lower risk of adverse effects. Several meloxicam formulations have been tested in horses, but a recently marketed granule oral formulation has not been studied. Objective: To characterize the pharmacokinetics of a novel granule meloxicam formulation in fasted and fed horses, and to compare pharmacokinetic features with oral suspension and tablets. Methods: Seven healthy adult horses. Methods: Meloxicam was administered at 0.6 mg/kg in fas...
Assessment of two methods to determine the relative contributions of the aerobic and anaerobic energy systems in racehorses. A prospective, randomized, controlled study was designed to determine relative aerobic and anaerobic (lactic and alactic) contributions at supramaximal exercise intensities using two different methods. Thoroughbred racehorses ( = 5) performed a maximal rate of oxygen consumption (V̇o) test and three supramaximal treadmill runs (105, 115, and 125% V̇o). Blood lactate concentration (BL) was measured at rest, every 15 s during runs, and 2, 5, 10, 20, 30, 40, 50, and 60 min postexercise. In , oxygen demand was calculated for each supramaximal intensity based on the V̇o test, and relative aerobi...
The lipidome of Thoroughbred racehorses before and after supramaximal exercise. A comprehensive study of the effect of supramaximal exercise in lipid homeostasis of Thoroughbreds provides the basis for future research on the role of lipids on energy metabolism in racehorses. Objective: To compare the plasma lipidome of Thoroughbreds before and after supramaximal exercise using an untargeted lipidomics approach. Methods: Pilot experimental study. Methods: Four Thoroughbred horses were used. The maximal oxygen consumption (VO ) was calculated for each horse. Horses then underwent treadmill exercise at the speed for which the oxygen requirements had been calculated to be 11...
Salivary alpha-amylase activity and concentration in horses with acute abdominal disease: Association with outcome. Salivary biomarkers could be useful to objectively evaluate critical illness and prognosis for survival in horses with acute abdominal disease. Objective: To compare salivary alpha-amylase (sAA) activity and concentration in healthy horses and horses with acute abdominal disease, and evaluate the association between sAA activity and concentration with disease severity and outcome. Methods: A prospective cohort. Methods: sAA activity, measured using a colorimetric commercial kit, and concentration, measured using a Time-resolved immunofluorometric assay, in 25 healthy horses and in 33 horses wi...
