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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.
Functional characterization of equine neutrophils in response to calcium ionophore A23187 and phorbol myristate acetate ex vivo. Equine neutrophils (PMN) play a critical role in inflammatory processes in horses. The objective of this study was to characterize equine PMN function ex vivo following stimulation with calcium ionophore A23187 (A23187) and phorbol myristate acetate (PMA). These stimulants trigger different branches of the PMN activation process that occurs in vivo. Equine PMN were isolated from the whole blood of six clinically normal geldings using a one-step discontinuous Percoll gradient technique. Neutrophil aggregation, degranulation, and superoxide anion production were evaluated in assay systems which ...
Purification and characterization of a hemolysin produced by Vibrio mimicus. Vibrio mimicus is a causative agent of human gastroenteritis. This pathogen secretes a pore-forming toxin, V. mimicus hemolysin (VMH), which causes hemolysis by three sequential steps: binding to an erythrocyte membrane, formation of a transmembrane pore, and disruption of the cell membrane. VMH with a molecular mass of 63 kDa was purified by ammonium sulfate precipitation and column chromatography with phenyl Sepharose HP and Superose 6 HR. The hemolytic reaction induced by VMH continued up to disruption of all erythrocytes in the assay system. Moreover, VMH that bound preliminarily to erythr...
The effects of ischemia and reperfusion on mucosal respiratory function, adenosine triphosphate, electrolyte, and water content in the ascending colon of ponies. The purpose of this study was to examine the effects of ischemia and reperfusion on the biochemical integrity of equine colonic mucosa to assess the relative roles of ischemic- and reperfusion-induced damage. Methods: Two hours of no-flow ischemia experimentally induced by 720 degrees counterclockwise ascending colon volvulus followed by 2 hours reperfusion after derotation. Methods: Ten ponies. Methods: Ascending colon biopsies were obtained every hour for measurement of mucosal adenosine triphosphate (ATP), water, sodium, and potassium content. Additional samples were homogenized for assay o...
The effect of strenuous versus moderate exercise on the metabolism of proteoglycans in articular cartilage from different weight-bearing regions of the equine third carpal bone. Articular cartilage degeneration in the middle carpal joint is a common problem in racing horses. This study evaluated the effect of exercise on the in-vitro synthesis of the large aggregating proteoglycans (aggrecan) and two small proteoglycans, biglycan and decorin, in articular cartilage taken from three weight bearing regions of the third carpal bone of horses which were subjected to moderate or strenuous exercise. Twelve Standardbred horses free from clinical and radiographic disease of the middle carpal joint were subjected to an 8 week moderate exercise program. The horses were then ran...
Role of endothelium and nitric oxide in the response of equine colonic arterial rings to vasoconstrictor agents. To determine the in vitro contractile responses of equine colonic arteries to angiotensin II, histamine, serotonin, norepinephrine, prostaglandin F2 alpha, vasopressin, and a thromboxane-B2-analogue. Methods: The tension generated in colonic arterial rings placed in organ baths with oxygenated Tyrode's solution at 37 degrees C after exposure to the previously mentioned chemical agents was measured using force-transducers interfaced with a polygraph. Methods: Large colon arterial rings collected from eight horses. Methods: The rings were allowed to equilibrate for 45 minutes after applying 2 g ...
Importance of sphingolipids and inhibitors of sphingolipid metabolism as components of animal diets. Sphingolipids are highly bioactive compounds that participate in the regulation of cell growth, differentiation, diverse cell functions, and apoptosis. They are present in both plant and animal foods in appreciable amounts, but little is known about their nutritional significance. Recent studies have shown that feeding sphingomyelin to female CF1 mice treated with a colon carcinogen (1,2-dimethylhydrazine) reduced the number of aberrant colonic crypt foci; longer-term feeding also affected the appearance of colonic adenocarcinomas. Therefore, dietary sphingolipids should be considered in studi...
Insulin-like growth factor 1 and corticosteroid modulation of chondrocyte metabolic and mitogenic activities in interleukin 1-conditioned equine cartilage. To evaluate potential stimulatory or matrix-sparing effects of insulin-like growth factor 1 (IGF-1), alone or in combination with a corticosteroid, in an interleukin 1 (IL-1)-induced model of cartilage degradation. Methods: Cartilage from the weightbearing surfaces of trochlea and condyles of clinically normal 2-year-old male horses. Methods: Triamcinolone acetonide and IGF-1 effects were evaluated by assessing: matrix responses by sulfated glycosaminoglycan (GAG) assay and [35S]sulfated GAG synthesis; collagen content by hydroxyproline assay; and mitogenic response by [3H]thymidine incorporat...
Distribution and functional effects of neuropeptide Y on equine ureteral smooth muscle and resistance arteries. The distribution of neuropeptide Y (NPY)-immunoreactive (IR) nerves, as well as the functional effects of NPY and the Y1- and Y2-receptor agonists, [Leu31,Pro34]NPY and NPY(13-36), respectively, have been investigated in vitro in both visceral and arterial smooth muscle of the horse intravesical ureter. NPY-IR nerve fibres were widely distributed along the entire length of the ureter, although the intravesical part was the most richly innervated region, and the only one where NPY-IR ganglion cells were found. NPY (10(-7) M) did not affect either basal tone or spontaneous rhythmic contractions ...
Isolation and characterization of heparin- and phosphorylcholine-binding proteins of boar and stallion seminal plasma. Primary structure of porcine pB1. In the bovine, seminal plasma heparin-binding proteins bind to sperm lipids containing the phosphorylcholine group and mediate the capacitating effects of heparin-like glycosaminoglycans during sperm residence in the female genital tract. We report the characterization of heparin- and phosphorylcholine-binding proteins of stallion and boar seminal plasma. Horse seminal plasma proteins HSP-1 and HSP-2, and boar protein pB1, belong to the same family as the bull heparin- and phosphorylcholine-binding proteins BSP-A1/2, BSP-A3, and BSP-30K. We have determined the amino acid sequence and posttrans...
Cloning, expression, purification, and characterization of the major core protein (p26) from equine infectious anemia virus. The gene coding for the major core protein (p26) of the lentivirus equine infectious anemia virus (EIAV) was cloned from EIAV infected serum, expressed in E. coli, and the resultant protein purified to electrophoretic homogeneity. The protein was expressed in a soluble form and was purified by conventional protein separation methods. When analyzed by SDS-PAGE, under both reducing and non-reducing conditions, the purified protein migrated as a 26 kDa monomer. Recombinant p26 (rp26), therefore, does not contain any intermolecular disulfide bond. Gel filtration chromatography also indicated that ...
In-vitro modulation of plasminogen activator activity, prostaglandin E and nitric oxide production by interleukin-1 in pregnant mare serum gonadotrophin-primed theca-interstitial cells. To examine the participation of the theca-interstitial (TI) compartment in cytokine modulation of ovarian function, the effects of interleukin-1beta (IL-1) on plasminogen activator (PA) activity and on prostaglandin E (PGE) and nitric oxide (NO) production were examined in cultures of pregnant mare serum gonadotrophin (PMSG)-primed rat TI cells. Exposure to IL-1 (10 ng/ml) resulted in a 25% reduction (P < 0.001) in PA activity, concurrent with a 4.6-fold increase in the ability of the corresponding conditioned media to inhibit exogenous urokinase activity. IL-1 also produced a 4.7-fold incr...
Biotransformation of chlorzoxazone by hepatic microsomes from humans and ten other mammalian species. The 6-hydroxylation of chlorzoxazone (CLZ) is currently being used in both in vivo and in vitro studies to quantify cytochrome P450 2E1 (CYP2E1) activity in humans. Comparatively little is known with regard to the biotransformation of this drug in other species. The NADPH-dependent biotransformation of CLZ was therefore studied using hepatic microsomes derived from humans and ten other mammalian species. In all species, 6-hydroxychlorzoxazone (6OH-CLZ) was the only metabolic product that could be identified by HPLC with ultraviolet detection. Enzyme kinetic analysis was used to characterize th...
Membrane contact with oviductal epithelium modulates the intracellular calcium concentration of equine spermatozoa in vitro. Interaction of equine spermatozoa with oviductal epithelial cells (OEC) prolongs sperm viability and maintains low intracellular calcium concentration ([Ca2+]i) in spermatozoa. Experiments were designed to investigate 1) whether release of spermatozoa from OEC in vitro is associated with elevated [Ca2+]i and 2) whether soluble products from OEC or direct membrane contact between spermatozoa and OEC mediates the effects of OEC on sperm [Ca2+]i. In the first experiment, changes in [Ca2+]i in spermatozoa loaded with indo-1 acetoxymethylester were determined in motile spermatozoa released from OEC...
Molten globule state of equine beta-lactoglobulin. The acid-unfolded state of equine beta-lactoglobulin was characterized by means of circular dichroism, nuclear magnetic resonance, analytical gel-filtration chromatography, and analytical centrifugation. The acid-unfolded state of equine beta-lactoglobulin has a substantial secondary structure as shown by the far-ultraviolet circular dichroism spectrum but lacks persistent tertiary packing of the side chains as indicated by the near-ultraviolet circular dichroism and nuclear magnetic resonance spectra. It is nearly as compact as the native conformation as shown by the gel filtration and sedime...
Prolactin, gonadotropin, and hair shedding responses to daily sulpiride administration in geldings in winter. This experiment was designed to determine 1) the efficacy of daily s.c. injections of a dopamine antagonist, sulpiride, for increasing prolactin secretion in geldings in winter and 2) whether increasing prolactin concentrations would hasten the onset of hair shedding or enhance gonadotropin secretion. Five geldings each received vehicle (vegetable oil) or sulpiride (100 mg in vehicle) daily from February 8 through March 29. On February 8 and every 7 d thereafter through March 29, blood samples were drawn around treatment injections and hair samples were collected. On March 30, all geldings rec...
Differential transcription of steroidogenic enzymes in the equine primary corpus luteum during diestrus and early pregnancy. In pregnant mares, eCG stimulates luteal androgen and estrogen production, increasing plasma concentrations 2- to 3-fold. To study how these changes are regulated, we examined the expression of mRNA for the steroidogenic enzymes 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD), cytochrome P450 17 alpha-hydroxylase/17,20-lyase (P450 17 alpha), and cytochrome P450 aromatase (P450arom) in equine primary corpora lutea using Northern blot analyses. Three equine specific cDNAs were generated by reverse transcriptase polymerase chain reaction. When compared to human, bovine, and rat sequences, the nu...
Localization of regulatory peptides in the male urogenital apparatus of domestic equidae: a comparative immunohistochemical study in Equus caballus and Equus asinus. An immunohistochemical study was carried out on specimens of testis, excurrent duct including the male accessory glands and urethra in its various tracts in the horse and the donkey, in order to localize nine regulatory peptides. Immunoreactivities were tested by means of Labelled Strept Avidin-Biotin (LSAB) method. The study has shown that Equine male genitalia are supplied by many peptide immunoreactive nerves containing NPY-, VIP-, leu- and met-Enkephalin-, Substance P-, CGRP- and Bombesin/GRP-like peptides, each of them having a characteristic distribution pattern. These neurotransmitters ...
Effect of maturation and aging on the histomorphometric and biochemical characteristics of equine superficial digital flexor tendon. To assess tendon morphology and non-reducible crosslink concentration, and associations of these findings with horse age and previously reported mechanical and ultrasonographic findings. Methods: Superficial digital flexor tendon samples were obtained from 23 horses aged 2 to 23 years. The tendons had undergone ultrasonography and were submitted to biomechanical testing in the physiologic range prior to sample acquisition. Methods: Samples were sectioned in a transverse plane; then dorsal, palmar, central, lateral, and medial regions were evaluated for fascicle cross-sectional area (CSA), sept...
Novel di-O-acetylated GM3s from equine erythrocytes, one containing 4,9-di-O-acetyl-N-glycolylneuraminic acid and another containing 4-O-acetyl-N-glycolylneuraminic acid and 6-O-acetyl-D-galactose. A novel GM3 O-acetylated at C-4 and at C-9 of N-glycolylneuraminic acid (4,9-di-O-Ac GM3), together with a second GM3 O-acetylated at O-4 of the neuraminic acid and O-6 of D-galactose (4,6'-di-O-Ac GM3) were isolated from equine erythrocytes as a mixture in approximate 1:1 ratio. These two major species were chromatographically inseparable. Their structures, especially the positions of the acetoxy group(s), were determined by means of 1D- and 2D-1H NMR and fast atom bombardment-MS as well as by gas chromatography-MS of partially O-methylated O-trimethylsilylated monosaccharides derived from th...
The hemoglobin polymorphism of the Sardinian wild dwarf horse and the oxygen binding properties of the four different horse hemoglobins. A study was made of the Hb phenotype of the Sardinian dwarfhorse (Equus caballus jara), one of the last surviving wild horse species in Europe. Hb haplotypes and their frequencies were found to be similar to those described in the Arabian horse (BI = 0.551, BII = 0.389, A = 0.036, V = 0.015), which suggests possible introduction onto the island from North Africa. The oxygen binding properties of the whole hemolysates and of the four different horse Hbs, separated by ion-exchange chromatography, were considered with regard to the effect of chloride, 2,3-bisphosphoglycerate and lactate. Results ...
Biochemical changes in the equine capsule following prostaglandin-induced pregnancy failure. The equine embryonic capsule, an acellular covering that envelops the conceptus during the second and third weeks of pregnancy, is composed of mucin-like glycoproteins. Its structure is consistent with a dual role during early pregnancy: protection of the conceptus, and communication between the embryo and the mother. Loss of sialic acid from the capsular glycoproteins at day 16 correlates with the time of "fixation," or loss of conceptus mobility throughout the uterine horns. This study investigated how the structure of the capsule is linked to the maintenance of pregnancy. Six pregnancies, c...
Oxidative energy metabolism in equine tendon cells. Hypoxia has been suggested as a possible cause of tissue degeneration and subsequent rupture in equine tendons. To determine whether low oxygen tension is likely to be detrimental to tendon cell function, experiments were designed to investigate oxidative energy metabolism in freshly isolated and cultured equine tendon cells. Freshly isolated tenocytes and cultured fibroblasts possessed activities of the mitochondrial enzyme citrate synthase similar to those of other mammalian cells, with well defined oxidative metabolism. D-[6(-14)C]-glucose oxidation was measurable in both freshly isolated a...
Methods for the isolation, culture and characterisation of equine pulmonary artery endothelial cells. Equine endothelial cells were isolated from the pulmonary artery by enzymatic digestion and grown to confluency. The cells were characterised by positive immunofluorescent staining for von Willebrand factor and NADPH-diaphorase staining for nitric oxide synthase. Measurements of endothelins indicated that there were significant release rates from the cells for up to six hours. Measurements of intracellular calcium concentration showed that the application of bradykinin caused a transient increase in calcium concentration with similar characteristics to those observed in other endothelial cell ...
Nitric oxide production by equine articular cells in vitro. Recent research in several species has suggested nitric oxide (NO) as a mediator of articular cartilage damage and an inhibitor of cartilage matrix neosynthesis. This study investigated NO production by cultured equine articular chondrocytes in response to 2 arthritogenic molecules, namely lipopolysaccharide (LPS) and interleukin-1 beta (IL-1 beta), and compared NO production by cultured equine synoviocytes stimulated with LPS. Synoviocytes exhibited a low basal level of NO synthesis (measured as nitrite, a NO metabolite) that was neither significantly increased nor decreased by exposure to LP...
Antibody directed against plasma membrane components of equine spermatozoa inhibits adhesion of spermatozoa to oviduct epithelial cells in vitro. Before fertilization, equine spermatozoa adhere to oviduct epithelial cells (OEC) of the mare. The biochemical basis for this adhesion has not been determined. Our objective was to produce an antiserum to block this interaction. Ejaculated spermatozoa were subjected to nitrogen cavitation and spermatozoal plasma membranes enriched by sucrose density gradient centrifugation; membrane enrichment was confirmed by comparative alkaline phosphatase analysis, electron microscopy, and one- and two-dimensional PAGE. Periacrosomal plasma membrane was used as an immunogen for the production of an antiser...
Fumonisins – Importance and occurence of a new group of mycotoxins. This paper describes the importance of fumonisins for human beings and animals and shows data for the occurence in food. Corn-based food samples (n = 299) purchased in the area of munich were analyzed for fumonisin content using an enzyme immunoassay.Fumonisins are mycotoxins produced byFusarium species, especially byFusarium moniliforme andFusarium proliferatum. Occurrence of fumonisins in corn and in cornbased foods and feeds has been reported from almost all over the world. In several animal species different diseases are traced back to fumonisin toxicosis. Fumonisin levels of 5-10 ppm inho...
Patterns of secretion of GnRH, LH and FSH during the postovulatory period in mares: mechanisms prolonging the LH surge. To study the mechanisms responsible for the unusually slow decline of the ovulatory LH surge in mares, secretion patterns of GnRH, LH and FSH were monitored in pituitary venous blood collected every 2 or 5 min for 10.5-18.0 h from five mares on the third (n = 4) or fifth day after ovulation (first sampling period). To determine the effectiveness of progesterone negative feedback, mares were then given a luteolytic dose of a prostaglandin analogue (PGF2 alpha) and pituitary venous sampling (every 2 or 5 min for 16 h) recommenced 20-22 h later (second sampling period). During the declining arm o...
Effect of feeding and feed deprivation on plasma concentrations of prolactin, insulin, growth hormone, and metabolites in horses. Two experiments were conducted to determine 1) the prolactin response to different kinds of feedstuffs in stallions and 2) the effects of total feed deprivation on prolactin secretion in mares and its interaction with the prolactin response to feeding. Experiment 1 was performed with stallions as a 6 x 6 Latin square: A) no feed; B) pelleted feed fed to meet 82.5% of the horses' CP requirements; C) pelleted feed at 25% of the amount in B; D) pelleted feed as in B plus water ad libitum; E) cracked corn at the weight in B; and F) chopped alfalfa at the weight in B. The positive prolactin respons...
A developmental switch in expression from blastocyst to endometrial/placental-type cytochrome P450 aromatase genes in the pig and horse. Pig blastocysts exhibit a transient period of estrogen production at periimplantation, with a second, more sustained period of estrogen synthesis occurring in endometrium and placenta at later pregnancy. Previously we reported the isolation of cDNA clones encoding a novel isoform of cytochrome P450 aromatase (the terminal enzyme in the estrogen biosynthetic pathway) from porcine periimplantation blastocysts. The present study investigated pregnancy-associated expression, in blastocysts and maternal reproductive tract tissues of this and an additional mRNA transcript encoding a distinct P450 ar...
Serum thyroid hormone concentrations in New Zealand horses. Total thyroxine and total tri-iodothyronine concentrations were measured in the sera from 125 horses of mixed age, breed and sex, and varied clinical histories. While low serum thyroxine concentrations were detected in 35 horses, the majority of those horses had serum thyroxine values within the reference range when retested. Only one horse had a mildly decreased serum tri-iodothyronine concentration. Those horses in which the serum thyroxine concentration was low when retested had a normal thyrotropin releasing hormone stimulation test. Hypothyroidism was not diagnosed in any horses in this s...
