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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.
Purification of horse muscle acylphosphatase antibodies by affinity chromatography. Horse muscle acylphosphatase antibodies were obtained by immunizing rabbits with the highly purified antigen cross-linked with glutaraldehyde. Specific antibodies were purified from the immunoglobulin fraction by affinity chromatography using a matrix coupled with the pure antigen as immunoadsorbent. The purified antibodies were partially characterized by immunodiffusion and immunoprecipitin techniques. These antibodies could be used to study aspects of the muscle acylphosphatase structure, localization and other biological properties.
[Purification of alpha-1,4 leads to 1,4-glucosyltransferase from horse blood serum]. The purification of alpha-1,4-1,4-glucosyltransferase from the equine serum is presented. Ion-exchange chromatography on DE-11, DE-32 and CM-32 celluloses was applied in the successive steps of isolation. Gel-filtration on Bio-Gel P-200 was the last step of purification; it gave the protein which was homogeneous on disc polyacrylamide gel electrophoresis. The purification degree was of the order 2100 at about 40% yield.
Isolation and identification of steroids from gonadal vein blood of the fetal horse. Direct connection of the artery of a fetal ovary to the carotid artery of the mare allowed collection of a large volume of blood over a 30-min period. Extraction of steroids and their fractionation was followed by separation of the steroids by alumina adsorption chromatography, and Sephadex LH-20 and Celite partition chromatography. Further resolution of the material by HPLC led to the identification of dehydroepiandrosterone (DHA) by nuclear magnetic resonance and mass spectrometry. Other compounds were isolated, which remain to be identified fully, but in the 8th month of pregnancy the princ...
[Properties of alpha-1,4 leads to -glucosyltransferase from the muscles and blood serum of horses]. Alpha-1,4 leads to 1,4-glucosyltransferase preparations from horse muscles and serum were studied. The enzyme proteins from both tissues are very similar. Both proteins have a molecular weight of 240 000 and consist of four subunits of 60 000 daltons each. pH 5,0 is optimal for the activity. Only substrates with alpha-1, 4-linkages can serve as glucosyl donors for transferase reactions. Km values for both enzymes differ very slightly. At low substrate concentrations the hydrolytic activity can be found in addition to transferase reactions. At the concentration of the substrate higher than 40 m...
Isolation and some properties of equine alpha 1-antitrypsin. 1. Equine alpha 1-antitrypsin was isolated from horse plasma by a combination of ammonium sulfate and acidification precipitation followed by ion-exchange chromatography on DEAE-cellulose, molecular sieve chromatography on Sephadex G-200 and affinity chromatography on Cibacron Blue-agarose. 2. The purified protein showed a single precipitin arc on immunoelectrophoresis in agarose but gave two bands on discontinuous polyacrylamide gel electrophoresis (PAGE). 3. Both bands appeared to interact equally with trypsin and were thought to represent two isoinhibitors of equine alpha 1-AT.
Subcellular distribution of particle-associated enzymes in horse neutrophil leukocytes. The subcellular components of purified neutrophil leukocytes from horse blood were fractionated by isopyknic equilibration in sucrose and metrizamide gradients. Five classes of particles have been identified: dense azurophil granules containing the bulk of the lysosomal acid hydrolase and peroxidase activity (A); less dense particles, containing all the lysozyme activity, but not resolved from a second population of azurophils B, and particles of low density, biochemically characterized as a plasma membrane fraction (C). Isopyknic equilibration in sucrose disclosed a minor membrane fraction (D...
Chemical composition of the spinal cord in the normal developing fetus and in the premature foal. The lipid content of spinal cord, expressed as a percentage of adult values, was considerably higher for newborn foals than for several other species and traces of esterified cholesterol (type A) were only rarely present in horse fetal cord (from 270 days gestational age onwards). This suggested that, at birth, the spinal cord is neurochemically more 'mature' in the horse than in cattle, sheep and pigs. Data for premature foals revealed no lipid abnormality suggestive of myelin immaturity or degeneration.
Energy metabolism in the erythrocytes of thoroughbred horses connected with perinatal physiological hemolysis. 1. The metabolism in the erythrocytes of thoroughbred horses in a sequential study from umbilical cord to the 1st month was investigated. 2. Emphasis was put on hemolytic period at which: (a). PFK, GSH-Px and GSH play a significant role. (b). There is a lower glucose consumption determined by a decreased activity in several enzymatic steps. (c). Singularly high concentrations of 2-3DPG and ATP were detected. 3. It has been suggested that the metabolic adjustments were achieved by an increased activity of the hexose monophosphate shunt, G-3PD and AK.
Interactions of different albumins and animal sera with insolubilized Cibacron Blue, Evaluation of apparent affinity constants. 1. A high concentration Cibacron Blue-Sepharose derivative has been used to study the affinity chromatography of albumin from eight animal species. 2. The apparent affinity constants for albumin varies between 3.9 x 10(4) M-1 and 0.9 x 10(4) M-1, in the order: Human greater than rabbit greater than horse greater than pig = dog greater than bovine greater than rat greater than chicken. 3. Other serum proteins were also bound to the gel, particularly lipoproteins and alpha 2-macroglobulin.
Primary structure of horse erythrocyte glycophorin HA. Its amino acid sequence has a unique homology with those of human and porcine erythrocyte glycophorins. The complete amino acid sequence of the major sialoglycoproteins of horse erythrocyte membranes, glycophorin HA, was determined by manual sequencing methods, using tryptic, chymotryptic, and cyanogen bromide fragments. Glycophorin HA is a polypeptide chain of 120 amino acid residues and contains 10 oligosaccharide units attached to the amino-terminal side of the molecule. Its amino terminus is pyroglutamic acid. All of the oligosaccharides are linked O-glycosidically to threonine or serine residues. The amino acid sequence is consistent with the transmembrane orientation of glycophorins. There...
The optimum pH of renal adenosine triphosphatase in rats: influence of vanadate, noradrenaline and potassium. In the presence of vanadate, the optimum pH of renal (Na+, K+)-ATPase in rats is reduced and lies in the range of intracellular pH. This explains the difference in optimum pH observed with ATP extracted from equine muscle. Removal of vanadate from such ATP (with noradrenaline) raises the optimum to the accepted range obtained with synthetic ATP. Changes in the sensitivity of the enzyme to potassium concentration contribute to the alterations in optimum pH. The optimum pH of Mg-ATPase is unaffected by vanadate. Since vanadate may be an intracellular regulator of (Na+, K+)-ATPase changes of opti...
Comparison of receptor properties of erythrocyte membrane glycoproteins. Membrane glycoproteins from horse, sheep, goat and bovine erythrocytes were solubilized and purified. These glycoproteins could be placed in three groups based on their degrees of glycosylation: The major bovine erythrocyte glycoprotein (BGII) had 77% sugar, the minor bovine glycoprotein (BGI) had 27% sugar and the others had approximately 50% sugar. Four of the glycoproteins aggregated in a uniform way in aqueous solution--one, BGII, did not. Four had similar subunit sizes of 25-34,000 daltons, but BGII was larger--55,000 daltons. Receptor functions (for plant and invertebrate lectins, antibo...
Effects of adenosine and deoxyadenosine on PHA-stimulation of lymphocytes of man, horse and pig. 1. Adenosine inhibits thymidine and uridine incorporation of PHA-stimulated lymphocytes of man and horse at concentrations higher than 50 and 10 microM, respectively. Deoxyadenosine is inhibitory at concentrations higher than 100 microM. Thymidine and uridine incorporation of porcine lymphocytes are elevated 5-7-fold by 25-100 microM adenosine, deoxyadenosine, inosine and hypoxanthine. Leucine incorporation of PHA-stimulated lymphocytes was affected by adenosine and deoxyadenosine in the same way, but to a lower extent. 2. Effects of adenosine and deoxyadenosine were more pronounced at shorter...
Identification of stage-specific and hormonally induced polypeptides in the uterine protein secretions of the mare during the oestrous cycle and pregnancy. Uterine secretions were obtained on Days 4, 8, 12, 14, 16, 18 and 20 of the oestrous cycle and early pregnancy. Acid phosphatase activity was significantly affected by day of the cycle, reaching a maximum at days 12-14 during the luteal phase and then declining to almost undetectable levels, by Day 20. In pregnant animals, activity continued to increase beyond Day 14. Two-dimensional polyacrylamide gel electrophoresis showed that albumin was a major component. However, a number of unique proteins of non-serum origin appeared in mid-cycle but had disappeared by Day 20. One of these was a basic ...
Radioimmunoassay and in-vitro bioassay of serum LH throughout the equine oestrous cycle. Mares were bled once daily throughout a cycle, or 3 times daily from the first day of oestrus to the 2nd day after ovulation. LH was measured by heterologous radioimmunoassay and by an in-vitro bioassay based on LH-stimulated testosterone production by mouse Leydig cells. The patterns of bio- and immuno-active LH during the oestrous cycle were similar but not identical, so that in both groups of mares the ratio of biological: immunological (B:I) activity during the LH surge was significantly higher before than after ovulation (P less than 0 . 001). Considerable individual variation in cycle me...
Vitamin A profiles of equine serum and milk. Serum and milk samples from mares and serum samples from their foals were taken at parturition and on d 1, 2, 4, 7, 14 and 21 postpartum. The samples were assayed for retinyl (r.) palmitate, r. acetate and retinol by high performance liquid chromatography. Peak vitamin A activity in milk occurred 1 d postpartum and preceded by 3 d the maximum vitamin A activity in foal serum and the lowest vitamin A activity in the mare serum. Mare serum contained approximately a 65:35 ratio of retinol:r. palmitate and less than 1% r. acetate. Retinyl palmitate was the predominant form of vitamin A in milk unt...
Effect of pregnancy and collection technique on prostaglandin F in the uterine lumen of Pony mares. Uterine flushings were obtained through the cervix (Method A) and through the wall of the uterus after hysterectomy (Method B) of ovariectomized Pony mares after s.c. injection of oestrogen for 1 week and progesterone for 2 weeks (Exp. 1). Non-pregnant and pregnant mares were flushed by Method A on Day 14 after ovulation and the flushings compared with those of non-pregnant mares injected i.v. with flunixen meglumine, a prostaglandin synthetase inhibitor, shortly before flushing (Exp. 3). Uterine flushings were also collected by Methods A and B from non-pregnant and pregnant Pony mares on Day ...
Equine immunology 2: immunopharmacology–biochemical basis of hypersensitivity. In general, 4 types of hypersensitivity reactions can be defined according to their immunological basis and clinical appearance. The differing mechanisms of these responses are described with particular reference to chemical mediators which through their pharmacological actions contribute to the clinical manifestations of hypersensitivity. Chemical mediators may exert their influence locally or systemically through their action on effector, tissues or organs and in addition, may be involved in the recruitment of cells of specific type to the site of the reaction. The possible role of these med...
Progesterone, prostaglandin F-2 alpha, PMSG and oestrone sulphate during early pregnancy in the mare. Blood samples from 4 mares during the late luteal phase, oestrus, early pregnancy and up to about 150 days of gestation were analysed for 15-keto-13,14-dihydroprostaglandin F-2 alpha (PGFM), progesterone, PMSG and oestrone sulphate by radioimmunoassays. During the late luteal phase, at the time of corpus luteum regression and decreasing progesterone levels, PGFM peaks were recorded. During early pregnancy (i.e. from mating and up to about Day 30) no such peaks were detected. After mating the progesterone levels increased and remained high throughout the observation period. During the oestrous ...
Changes in equine follicular aromatase activity during transition from winter anoestrus. Follicular aromatase activity during sexual resurgence after the winter anoestrus was investigated in 3 groups of 5 Pony mares. Group ET was studied during the early transition period, Group LT in late transition and Group C in full breeding condition. Granulosa and theca cells were incubated for 3 h with 3H-labelled androstenedione or progesterone. Analysis of the free oestrogenic products was by high performance liquid chromatography (HPLC) and recrystallization revealed highly variable oestrogen production in both cell types from mares in all 3 groups. Only oestrone and oestradiol peaks wer...
Furosemide, Patella vulgata beta-glucuronidase and drug analysis: conditions for enhancement of the TLC detection of apomorphine, butorphanol, hydromorphone, nalbuphine, oxymorphone and pentazocine in equine urine. We have investigated the action of five sources of beta-glucuronidase enzymes on the hydrolysis of glucuronides of apomorphine, butorphanol, hydromorphone, nalbuphine, oxymorphone and pentazocine in equine urine. For all glucuronides tested, Patella vulgata beta-glucuronidase yielded the largest thin layer chromatographic (TLC) spots. For oxymorphone, P. vulgata was the only treatment to yield detectable TLC spots under test parameters. For these six drugs, TLC spot size and chromatographic quality were compared between control horses and horses pretreated with furosemide four hours earlier. F...
Changes in plasma progesterone levels during storage of heparinized whole blood from cow, horse, dog and pig. Progesterone concentrations in heparinized plasma harvested immediately after blood collection were compared with levels obtained after storage of the corresponding whole blood for 2 h, 4 h, 6 h, 1 day, 2 days and 5 days at room temperature and in a refrigerator. The blood was taken during the luteal phase from 4 dogs, 4 horses, 4 pigs and 8 cows. For 4 cows the storage time was extended to 9 and 20 days. No significant effect of whole blood storage time on plasma progesterone concentrations could be shown for dogs or pigs. For the horse a slight but significant decrease was demonstrated when ...
Dehydroepiandrosterone synthesis by the fetal foal and its importance as an oestrogen precursor. The gonads of the fetal horse were found to be relatively devoid of 3 beta-hydroxysteroid dehydrogenase and other enzymes which metabolize dehydroepiandrosterone (DHA). In short-term in-vitro incubation experiments fetal liver converted DHA to the potential equilin precursor, 7 alpha-hydroxy DHA. DHA was converted to oestrone when incubated with extracts of horse placenta but 7 alpha-hydroxy DHA was not converted to equilin. Levels of DHA measured in peripheral blood of mares throughout pregnancy paralleled those of equilin and oestrone, and DHA concentrations fell rapidly after fetal gonadect...
Comparison of the interaction of equine LH and human chorionic gonadotrophin to equine testicular receptors. Human chorionic gonadotrophin (hCG) can be used to study horse luteinizing hormone (LH) receptors in stallion testicular tissue. hCG was more stable than horse LH during radioiodination when compared by their abilities to bind to testicular receptor sites. During incubation, neither hormone lost binding activity at 4 degrees C. Horse LH lost binding activity during incubation at 25 degrees C and both hormones lost binding activity at 37 degrees C. Both hormones bound to the same receptor sites which are specific for the hormones. The receptor sites were not degraded when incubated at 4 degrees...
Effect of PGF-2 alpha on LH receptors in the equine corpus luteum. As quantified by Scatchard analysis, a 27 000 g crude luteal membrane fraction contained a single population of unoccupied LH receptors characterized by high affinity, ka = 0.647 +/- 0.158 X 10(11) M-1 and low binding capacity, Rt = 4.91 +/- 0.78 X 10(-11) M/mg membrane fraction. Acceptable hormonal specificity, reversibility, saturability, high affinity and tissue specificity indicated that the binding protein was a physiological receptor. To ensure that the methods used for Scatchard analysis were valid, hCG was characterized for specific activity and maximum bindability, non-specific bindin...
Titration of antiserum to South American rattlesnake (Crotalus durissus terrificus) venom by measuring inhibition of phospholipase A2 activity. Horse antiserum to the venom of Crotalus durissus terrificus, A South American rattlesnake, inhibits the phospholipase activity of the crude venom. There is a close relationship between this inhibitory property and the neutralizing potency of the antiserum in vivo. This may provide the basis for a rigorous standardization of anticrotalid venom in vitro.
