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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.
Localization of xanthine dehydrogenase mRNA in horse skeletal muscle by in situ hybridization with digoxigenin-labelled probe. In situ hybridization was used to localize xanthine dehydrogenase (XDH) mRNA in horse skeletal muscle. Capillary endothelial cells were found to express XDH, but muscle cells did not give any signal. The digoxigenin-labelled probe was produced by PCR with primers based on the cDNA sequence of mouse XDH and horse lung cDNAs. A 4.3 kb mRNA was detected in a Northern blot.
The effect of cortisol on the secretion of ACTH by anterior pituitary cells of the horse in culture. Perifused equine anterior pituitary cells were used to investigate the effect of cortisol on the ACTH response to pulses of corticotrophin-releasing hormone (CRH; 0.01 nmol/l) and arginine vasopressin (AVP; 100 nmol/l), given for 5 min every 30 min for 690 min and ACTH measured in 5-min fractions. At the fourth pulse of secretagogue (0 min), a constant perifusion with cortisol began (0 nmol/l (control), 100, 200, 500, 5000 and 50,000 nmol/l) and continued until the ninth pulse (150 min). For each pulse of secretagogue, the amount of ACTH (pmol) secreted in response to each pulse (ACTH response...
Placental and plasma cystine aminopeptidase in pregnant animals. The placental and plasma cystine aminopeptidase (CAP) in pregnant animals was examined on stability after the treatment with L-methionine, ethylene diamine tetra-acetic acid (EDTA) and heat. Inhibitory effects of these treatments on enzyme activities were different among CAPs from the animal species, however, significant correlation in those effects between placental and plasma CAPs was observed. These results suggested that plasma CAP might reflect placental CAP and seemed to be available for estimating maternal gestational conditions.
Isolation of an inhibitor of tumor necrosis factor-alpha-mediated cytotoxicity liberated from chemotaxin-stimulated equine white blood cell populations. Objectives of this investigation were to extract and isolate protein fractions inhibitory to the cytotoxic properties of tumor necrosis factor-alpha (TNF-alpha). In this context, mixed populations of WBC were harvested from equine blood and were stimulated with a combination of a synthetic chemotactic peptide and a calcium ionophore. Several methods were subsequently applied for the initial preparation of cell-free crude protein extracts, including fractional precipitation with gradient concentrations of ammonium sulfate and preparative-scale isoelectric focusing. In addition, protein fraction...
In vitro concentrative accumulation of D-xylose by jejunum from horses and rabbits. Accumulation of D-xylose by jejunal mucosa from healthy horses and rabbits was studied in vitro. When tissue sheets were incubated with 1 mM D-xylose for 60 minutes, mucosa from horses and rabbits accumulated D-xylose against a concentration gradient. There was no accumulation when equine specimens were incubated with 5 mM D-xylose. By comparison, equine jejunum accumulated D-glucose against a concentration gradient when incubated in 5 mM D-glucose. In equine and rabbit jejunum, 13.3 +/- 7.0% and 36 +/- 11.0%, respectively, of accumulated D-xylose was phosphorylated when sheets were incubated ...
Neuropeptide distributions in the colon, cecum, and jejunum of the horse. The pelvic flexure portion of the equine large colon is the proposed location of a pacemaker mechanism. This study was conducted to ascertain whether the distribution of certain putative neurotransmitters differs at the pelvic flexure compared to other sampling sites. Tissue samples were collected from the intestinal tracts of six horses. Serial sections from these samples were reacted with primary antisera specific for substance P, vasoactive intestinal polypeptide (VIP), methionine-Enkephalin, and calcitonin gene-related peptide (CGRP). The regional distribution of immunoreactive neuronal el...
Different in vitro metabolism of 7 alpha-methyl-19-nortestosterone by human and equine aromatases. The ability of human and equine placental microsomes to aromatize 7 alpha-methyl-19-nortestosterone (MNT) was studied. Kinetic analysis indicates that MNT shares the androgen-binding site of human and equine placental microsomal aromatases. Human placental microsomal estrogen synthetase had about a 2.5-fold higher relative affinity for MNT than the equine placental enzyme (KiMNT/Km androstenedione of 32 versus 87). However, MNT was not metabolized by human placental microsomes, whereas it was very actively metabolized by equine placental microsomes. Further studies using purified equine cytoch...
Immunocytochemical localization of some turkey pituitary hormones using antisera to human hormones. This study was conducted to determine the crossreactivity of antisera to human prolactin (PRL), adrenocorticotropic hormone (ACTH), and growth hormone (GH) to turkey pituicytes. In addition, crossreactivities of the above antisera and antiserum to turkey GH to pituicytes of turkey, cat, rabbit, horse, owl monkey, and human were evaluated. Results of the immunocytochemical localizations showed that with one exception antisera to human hormones were positive for each species tested. Turkey pituicytes failed to crossreact with antiserum to human GH. Likewise, antiserum to turkey GH failed to cros...
Developmental regulation of insulin like growth factor II expression in the horse. The expression of the insulin like growth factor (IGF) II gene has been examined in the developing equine fetus. It was found that IGF II transcripts were present in abundant quantities in third trimester embryonic and extraembryonic tissues as for example the placenta. The expression of the IGF II gene was high in the fetal liver where two prominent transcripts--4.6 and 4.1--kB were produced. However, these transcripts could not be traced in the adult liver. Instead we found two different transcripts with the sizes of 4.0 and 2.9 kB in the adult liver. These findings taken together with the d...
Factors affecting ACTH release from perifused equine anterior pituitary cells. The multifactorial control of ACTH is well established. We wished to establish and characterize an in-vitro perifusion system, using equine anterior pituitary cells and physiological concentrations of secretagogues, to investigate factors which affect the dynamics of ACTH secretion. Anterior pituitary tissue was divided for dispersion into cells with collagenase, trypsin or dispase, or by mechanical dispersion. After dispersal followed by 18-h incubation, cells were perifused and the ACTH response to 10-min pulses of arginine vasopressin (AVP; 100 nmol/l), corticotrophin-releasing hormone (CRH...
Structural relaxation and nonexponential kinetics of CO-binding to horse myoglobin. Multiple flash photolysis experiments. The geminate recombination kinetics of CO-myoglobin strongly deviates from single exponential behavior in contrast to what is expected for unimolecular reactions (1). At low temperatures, this result was attributed to slowly exchanging conformational states which differ substantially in barrier height for ligand binding. Above 160 K the kinetics apparently slow down with temperature increase. Agmon and Hopfield (2) explain this result in terms of structural relaxation perpendicular to the reaction coordinate, which enhances the activation energy. In their model, structural relaxation homogeniz...
A liquid chromatographic procedure for the analysis of yohimbine in equine serum and urine. A standardbred mare was dosed with 40 mg yohimbine intravenously. Serum and urine samples were collected and analyzed for yohimbine using solvent extraction and reversed-phase high-performance liquid chromatography (HPLC) with fluorescence detection. Maximum yohimbine concentrations of 45 and 18 ng/mL were observed in serum and urine samples, respectively. Elimination was rapid, with half-lives of approximately 20 and 53 min observed for serum and urine, respectively. The presence of yohimbine in these samples was confirmed by liquid chromatography/mass spectroscopy (LC/MS/MS).
Plasma constituents during incremental treadmill exercise in intact and splenectomised horses. Six intact and 6 splenectomised mares were given an incremental exercise test on a treadmill to examine the fluid and electrolyte changes associated with exercise and the role of the spleen in these changes. Blood samples were obtained at rest and at the end of each 1-min step of the test. Exercise at 7 m/sec caused significant (P 0.05) in either group (intact = 2.7%; splenectomised, = 3.5%). This appears to be the first record of substantial changes of these constituents during short-term exercise, even before the onset of visible sweat losses. The changes in the concentration of plasma prot...
Preliminary investigations into factors that affect plasma aldosterone concentrations in horses. The effect of feeding diets with low, adequate and high sodium contents on plasma aldosterone concentrations in horses and ponies was evaluated using human immunoassay kits. The effect of moderate to high intensity exercise of up to six minutes duration on plasma aldosterone concentrations in three thoroughbred horses was also investigated. On an adequate sodium diet plasma aldosterone concentrations increased to a peak around four hours after feeding. Little daily variation was found in the pre-feeding aldosterone concentrations over three days. Feeding additional salt resulted initially in n...
Equine ovarian aromatase: evidence for a species specificity. Mare granulosa cells and cyclic corpus luteum microsomes are reported to aromatize 19-norandrogens more efficiently than androgens. However, 16 alpha-hydroxytestosterone and epitestosterone were not aromatized by the equine corpus luteum microsomal estrogen synthetase. These results indicate that the equine aromatase system would be different from the human placental microsomal estrogen synthetase, which aromatizes 16 alpha-hydroxyandrogens and epitestosterone but not 19-norandrogens. Furthermore, our data show that the rates of aromatization of androgens and 19-norandrogens were not additive ...
Catecholaminergic innervation of the equine ureter. The aim of the present study was to determine the distribution and density of catecholaminergic nerve fibres and cells in the equine ureter by using immunohistochemical techniques to localise the enzyme tyrosine-hydroxylase (TH). TH-immunoreactive (TH-IR) nerve fibres entered the wall of the ureter as adventitial nerve trunks accompanying the blood vessels. These trunks repeatedly branched as they coursed through the muscular layer towards the epithelium, forming muscular, perivascular and subepithelial nerve plexuses. TH-IR nerve fibres were especially numerous in the pelvic and intravesical ...
Focal exocytosis by eosinophils–compound exocytosis and cumulative fusion. We have investigated the granule fusion events during exocytosis in horse eosinophils by time-resolved patch-clamp capacitance measurements. Stimulation with intracellular GTP gamma S leads to a stepwise capacitance increase by 4.0 +/- 0.9 pF. At GTP gamma S concentrations < 20 microM the step size distribution is in agreement with the granule size distribution in resting cells. Above 80 microM the number of steps is reduced and very large steps occur. The total capacitance increase, however, is unaffected. These results show that at high GTP gamma S concentrations granule--granule fusion o...
Production and characterization of a monoclonal antibody recognizing a cytoplasmic antigen of equine mononuclear phagocytes. An IgG1 mouse monoclonal antibody, designated 1.646, is described which recognizes a cytoplasmic antigen of equine mononuclear phagocytes. Indirect fluorescent antibody staining of peripheral blood leukocytes reveals a granular cytoplasmic staining, predominantly in adherent blood mononuclear cells. Indirect fluorescent antibody staining is positive for alveolar and peritoneal macrophages. In some horses, a few neutrophils are also stained. In equine tissue samples stained by immunohistochemistry, the distribution of positive cells is consistent with the distribution of tissue macrophages. The...
Isolation and characterization of four basic proteins from horse eosinophilic granules. Four new basic proteins were isolated from horse eosinophils and purified. The eosinophils release these proteins after permeabilization with saponin and degranulation stimulized by guanosine 5'-O-thiotriphosphate. The proteins were separated and purified on a Superose P12- and a Mono S-column by fast protein liquid chromatography. The amino acid composition, the relative molecular mass, the isoelectric point and the partial N-terminal sequence of the four proteins were determined. Papain-activation and ribonuclease activity of the four proteins were tested for comparison with the human eosino...
Competitive inhibition of lipolytic enzymes. IX. A comparative study on the inhibition of pancreatic phospholipases A2 from different sources by (R)-2-acylamino phospholipid analogues. The inhibitory power (Z) of a number of (R)-1-alkyl-2-acylamino phospholipid analogues was determined for three mammalian phospholipases A2 from pig, ox and horse pancreas. All three enzymes display a clear preference for anionic (phosphoglycol) inhibitors over the zwitterionic (phosphocholine) derivatives; this effect is most pronounced for the bovine enzyme. Upon variation of the 1-alkyl chain length, the bovine and equine phospholipases, like the porcine enzyme in previous studies, show an optimum in Z for a six-carbon alkyl group. The introduction of a double bond in the 2-acylamino group ...
Immunoaffinity chromatography combined with gas chromatography-negative ion chemical ionisation mass spectrometry for the confirmation of flumethasone abuse in the equine. Immunoaffinity chromatography using a synthesised immunosorbent was used to extract tritiated dexamethasone (with dexamethasone carrier) from equine urine at a recovery of 81.7 +/- 8.4% (mean +/- S.D.). A method utilising this procedure coupled to cool on-column injection gas chromatography-negative ion chemical ionisation mass spectrometry is also described for the confirmation of low levels of flumethasone in equine urine samples.
Studies on the substrate specificity of the proteinase of equine infectious anemia virus using oligopeptide substrates. The proteinase of the equine infectious anemia virus (EIAV), a lentivirus closely related to human immunodeficiency virus (HIV), was purified from concentrated virus. The specificity of the enzyme was characterized using oligopeptides representing naturally occurring cleavage sites in the Gag and Gag-Pol polyproteins. The length of the substrate binding pocket was found to be 1-2 residues longer than that of HIV proteinases. Although the EIAV and HIV proteinases cleaved most of the peptides at the same bond, some were hydrolyzed by only the EIAV enzyme. Oligopeptides representing cleavage site...
Comparative immunohistolocalization of carbonic anhydrase isozymes I, II and III in the equine and bovine digestive tract. Immunohistochemical localizations of carbonic anhydrase isozymes (CA-I, CA-II and CA-III) in equine and bovine digestive tracts were studied. In the horse, epithelial cells in both the oesophagus and non-glandular part of the stomach lacked all three isozymes. In contrast, surface epithelial and parietal cells in the glandular region of the stomach showed reactivity for CA-II. In the small intestine, absorptive columnar cells covering the villi in the duodenum were positive for CA-II. The epithelium of the jejunum and ileum lacked all three isozymes. In the large intestine, CA-II was detected ...
Characterization of serum lysosomal enzymatic activities. III. Effect of infectious influenza in Egyptian equines. An outbreak of infectious influenza was recognized in Menofeia governorate in October 1989. Eight naturally influenza infected as well as 8 healthy control horses, mules and donkeys were selected for collection of blood and sera separation to estimate four lysosomal enzymatic activities and to describe the clinical findings, which were fever, congested nasal, conjunctival membranes and cough. Bronchopneumonia followed later with bilateral purulent nasal discharge as a complication in 2 donkeys. Thereafter laboured breathing occurred. Therefore a therapeutic penicillin-streptomycin dose was inj...
Distribution of dopamine beta-hydroxylase and neuropeptide Y-immunoreactive nerves in healthy equine lungs. Immunohistochemical methods were used to determine the distribution of pulmonary nerves containing either an enzymatic marker of adrenergic nerves, dopamine beta hydroxylase, or the putative neurotransmitter neuropeptide Y in 7 equids with healthy lungs. Nerves immunoreactive for these substances were found on airway smooth muscle in nearly all the samples of healthy equine lung examined. These nerves were generally more numerous in the larger airways but could be detected even in noncartilaginous bronchioles. Pulmonary and bronchial vessels also contained numerous immunoreactive nerves. On th...
A novel response of anion transporter in equine erythrocytes to a fluorescent substrate, N-(2-aminoethyl sulfonate)-7-nitrobenz-2-oxa-3-diazole (NBD-taurine). This report describes a unique response of the anion transporter in equine erythrocytes to the fluorescent substrate N-(2-aminoethyl sulfonate)-7-nitrobenz-2-oxa-3-diazole (NBD-taurine). Equine erythrocytes showed fluxes of NBD-taurine both inward and outward at rates considerably slower than those in human cells. These fluxes were completely abolished by a typical anion transport inhibitor, 4,4'-diisothiocyanostilbene-2,2'-disulfonate. Furthermore, NBD-taurine competitively inhibited the uptake of phosphate in equine red cells with an inhibition constant of phosphate that was slightly higher ...
Topography of equine chorionic gonadotropin epitopes relative to the luteinizing hormone and follicle-stimulating hormone receptor interaction sites. In order to localize the epitopes of equine chorionic gonadotropin (eCG) involved in interaction with luteinizing hormone (LH) and follicle-stimulating hormone (FSH) receptors, we used 14 monoclonal anti-eCG antibodies (mAbs). Different effects of these mAbs on the bioactivities of eCG were observed in in vitro bioassays, but the effects of each mAb on the two bioactivities were similar for all but four mAbs. All mAbs were found to inhibit the binding of eCG to LH receptors except 3A3 mAb, in radioreceptor assay. Six mAbs, which were strong inhibitors of eCG binding to LH receptors and of both...
Echinocytosis in horses: 54 cases (1990). Retrospective review of CBC and serum chemical data from 124 horses admitted to the veterinary teaching hospital over a 9-month period (Feb 1, 1990 to Oct 31, 1990) indicated that 54 horses had echinocytosis (prevalence = 44%). In horses with echinocytosis, the most frequent diagnosis was colitis (23 horses; 43%). Odds ratios (measure of association) were calculated to determine the association of echinocytosis with specific hematologic and biochemical abnormalities. When evaluated in a multivariate model, low serum sodium concentration (< 136 mEq/L) was the only variable significantly associa...
