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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.
Exercise-induced phospholipid degradation in the equine skeletal muscle and erythrocytes. To understand the pathogenesis of equine exercise-induced myopathies and hemolysis, changes of phospholipid peroxidation products in the equine middle gluteal muscle and erythrocytes following the high-speed treadmill exercise were studied. In the skeletal muscle, the peroxidized phosphatidylethanolamine (PE) were increased at 24 hours after the exercise. The malondialdehydes (MDAs) were also increased as the protein-bound MDAs following exercise. In the erythrocytes, the peroxidized PE were significantly decreased at 24 hours after the exercise. The protein-bound MDAs were significantly incre...
Cloning and characterization of gene TNF alpha encoding equine tumor necrosis factor alpha. We report the molecular cloning and nucleotide sequence of the equine gene encoding tumor necrosis factor alpha. The 2610-bp genomic sequence was derived from three overlapping polymerase chain reaction products.
Purification and kinetic characterization of equine infectious anemia virus reverse transcriptase. The reverse transcriptase of Equine Infectious Anemia Virus (EIAV) was partially purified from virus particles and appeared to be a heterodimer with subunit molecular masses of 70 kdal and 59 kdal. The polymerase activity of this enzyme had an absolute requirement for a divalent cation, preferring Mg++ over Mn++. Addition of a monovalent cation to the reaction mixture enhanced, but was not required for enzyme activity. Kinetically, the reverse transcriptase of EIAV is similar to the reverse transcriptase of Human Imunodeficiency Virus Type 1 (HIV-1). Both enzymes have similar Km values for 2'-...
Structure of equine corticotropin releasing factor. A 41 amino acid peptide, probably identical in structure to human corticotropin releasing factor, was isolated from 70 equine hypothalami by methanol extraction, immunoaffinity chromatography and single step of reverse phase HPLC. The amino acid sequence was determined by gas phase sequence analysis. Probable carboxyl terminal amidation was demonstrated by similar retention times for equine and human corticotropin releasing factor on reverse phase HPLC at pH 8. The likely structure of equine corticotropin releasing factor is: Ser-Glu-Glu-Pro-Pro- Ile-Ser-Leu-Asp-Leu-Thr-Phe-His-Leu-Leu-Arg-Glu...
Plasma histamine levels in laminitic horses and in horses treated with a corticosteroid. In seven sound horses histamine plasma levels were evaluated by means of a radioenzymatic method using the enzyme histamine-N-methyltransferase (mean = 4.46 nmol/l, SD = 1.55 nmol/l). Histamine levels of laminitic horses were significantly higher (n = 11; mean = 12.46 nmol/l, SD = 3.24 nmol/l). In three sound horses plasma histamine levels were determined over a two week period following a double administration of 10 mg dexamethasone-21-isonicotinate/100 kg body weight. Histamine levels remained within the range of sound control horses.
Effect of storage on measurement of ionized calcium and acid-base variables in equine, bovine, ovine, and canine venous blood. The stability of blood ionized calcium (Ca2+) and acid-base variables in equine, bovine, ovine, and canine venous blood samples (n = 15, in each group) stored at 4 C for 3, 6, 9, 24, or 48 hours was studied. Variables included blood Ca2+ and standard ionized calcium (Ca2+ corrected to pH 7.4) concentrations, pH, blood carbon dioxide and oxygen tensions, base excess, bicarbonate concentration, and total carbon dioxide content. Results indicate that storage of blood samples at 4 C for up to 48 hours, despite appreciable acid-base changes, is associated with less than 1.5% change in equine, bovin...
Measurement of oxytocin concentrations in plasma and ovarian extracts during the oestrous cycle of mares. Jugular venous blood samples were collected throughout a complete oestrous cycle from 9 mares for measurement of progesterone and oxytocin by radioimmunoassay. Mean oxytocin concentrations remained at approximately 1 pg/ml throughout, with no evidence of cyclic variation in the release pattern. Extracts of corpus luteum and follicles obtained from a further 33 mares at different stages of the cycle all contained oxytocin concentrations of less than 10 pg/g wet weight of tissue. We conclude that the ovaries are not a source of circulating oxytocin during the oestrous cycle in this species. The ...
Stability of equine lysozyme. I. Thermal unfolding behaviour. The thermal denaturation of Ca(2+)- and apo-forms of equine lysozyme was followed by using far and near UV circular dichroism and intrinsic fluorescence methods. The difference found between the temperature dependence of the ellipticity at 222 nm and 287 nm, which show two stages in the thermal transition, and those at 228 nm and 294 nm, which indicate only one stage over a wide range of temperatures reflects that different subdivisions of the protein molecule are characterized by a different stability, cooperativity and pathway of denaturation. The first transition, reflected in the increase ...
Value of standardised exercise tests and blood biochemistry in the selection and training of breeding stallions. Stallions selected by the Royal Dutch Warmblood Society were submitted to a standardised lungeing test at the beginning and at the end of a 100-day test of performance and ability. The heart rate, haematology and biochemistry values obtained in the first lungeing test showed no significant differences between the 15 stallions which were rejected by the Royal Dutch Warmblood Society during the first month of the 100-day test, the 15 stallions rejected during the last month and the 11 stallions which were approved for registration in the studbook. The 26 stallions submitted to the second lungein...
Prostaglandin E2 secretion by oviductal transport-stage equine embryos. This study was conducted to identify embryonic products whose secretion was temporally associated with the oviductal transport period of the mare. Chemicals secreted by oviductal-transport-stage equine embryos were identified by incubating Day 6 or Day 7 early uterine embryos with 35S-methionine/cysteine, 3H-progesterone, or 3H-arachidonic acid for 24 h, and subsequently identifying radioactively labeled proteins (SDS-PAGE; n = 3 embryos), steroids (HPLC; n = 3 embryos), or prostaglandins (HPLC; n = 3 embryos) in the culture medium. Early uterine embryos secreted 116.1 +/- 45.5 pg of prostagla...
Metabolic responses to ammonium acetate infusion in exercising horses. The relationship between elevated plasma ammonia (NH3) levels, fatigue development and muscle metabolism were examined in horses during a submaximal fatigue test. Eight Quarter Horse mares were intravenously infused prior to exercise with either sodium acetate (control) or ammonium acetate (AMINF), and exercised to fatigue on an 11% grade treadmill, carrying 27 kg of lead. Time to fatigue was not different (P greater than 0.05) between groups. Intramuscular NH3 and lactate increased (P less than 0.001) during exercise; however, the treatment did not (P greater than 0.05) affect either. A treat...
The equine major plasma serpin multigene family: partial characterization including sequence of the reactive-site regions. The equine Pi system, which is highly polymorphic and was considered to be controlled by a single locus, has been shown to be controlled by four loci (named Spi 1-4). This system is the equine equivalent of the major human plasma serpin (serine protease inhibitor), human alpha 1 PI. Twenty-two haplotypes of the equine Pi system have been characterized by two-dimensional electrophoresis, resulting in the assignment of pI, Mr, and bovine trypsin and chymotrypsin inhibition characteristics to 109 proteins. These proteins have been analyzed further to determine their relatedness to each other as w...
Xanthine oxidase formation during experimental ischemia of the equine small intestine. We hypothesized that xanthine oxidase plays a role in the postischemic reperfusion injury in the equine small intestine. Under anesthesia, four horses and two ponies underwent ischemic strangulating obstructions of segments of the proximal jejunum, mid-jejunum and ileum. Prior to vascular occlusion, and at 1 h and 2 h of ischemia, full-thickness intestinal biopsies were collected for histopathological evaluation and for determination of combined xanthine dehydrogenase (XDH) plus xanthine oxidase (XO) activity, and XO activity alone. The level of XO activity was expressed in percentage accordin...
Changes of plasma concentrations of steroid hormones, prostaglandin F2 alpha-metabolite and pregnant mare serum gonadotropin during pregnancy in thoroughbred mares. Plasma concentrations of estrogens, gestagens, cortisol (F), 13, 14-dihydro, 15-keto PGF2 alpha (PGFM) and pregnant mare serum gonadotropin (PMSG) in 10 Thoroughbred mares were measured for a 11-month pregnancy period. Estrone (E1) and estradiol-17 beta (E2) levels gradually increased as the pregnancy advanced, showing a peak around Month 8 and decreased thereafter. Progesterone (P) levels increased on Months 3 and 11, and 17 alpha-OH-progesterone (17 alpha-OHP) levels peaked on Month 3, whereas 20 alpha-OH-progesterone (20 alpha-OHP) levels increased sharply after Month 6. PGFM indicated peak...
Effects of carbohydrates on the pharmacokinetics and biological activity of equine chorionic gonadotropin in vivo. The sialylation of eCG was examined to determine its influence on the in vivo metabolism and biological activity of the molecule. Sialic acid was decrementally removed from eCG by incubation with agarose-linked neuraminidase for varying time periods. Pharmacokinetic parameters for the disappearance of 4,000 IU (267 micrograms) of three desialylated eCG preparations (20%, 53%, and 80% sialic acid removed) and control eCG were determined in sheep. The clearance rate of eCG increased (p less than 0.05) with each decrement of sialic acid. The removal of 53% sialic acid enhanced the distribution of...
Participation of H1-receptors in histamine-induced contraction and relaxation of horse coronary artery in vitro. The mechanisms of histamine-induced contraction and relaxation were investigated in rings isolated from a middle part of the left descending coronary arteries of horses. Intact and endothelium-denuded preparations were compared. Rings of horse coronary arteries contracted in response to histamine in a concentration dependent manner, but some of them relaxed with lower concentrations and contracted with higher concentrations. Removal of the endothelium abolished the relaxation and potentiated the contraction. The pD2 values were 4.70 +/- 0.08 in the rings with intact endothelium and 4.95 +/- 0....
Influence of polysulfated glycosaminoglycan on equine articular cartilage in explant culture. Articular cartilage explants from 3 horses were maintained in tissue culture to test the effects of a polysulfated glycosaminoglycan on proteoglycan biosynthesis. Cultures were exposed to concentrations of 0, 50, or 200 micrograms of the drug/ml for either 2 days or 6 days, and labeled with 35S, before measuring the content of sulfated proteoglycan in the culture media and in extracts of cartilage. In a second experiment, the explants were incubated with the isotope and subsequently exposed to the same concentrations of the polysulfated glycosaminoglycan for 4 days. Subsequently, the amount of...
Effect of whole-body potassium depletion on plasma, erythrocyte, and middle gluteal muscle potassium concentration of healthy, adult horses. The effects of whole-body potassium depletion induced by food deprivation on plasma, erythrocyte, and middle gluteal muscle K concentrations was quantified in 16 healthy, adult horses before, during, and at the end of a 7-day period of food deprivation during which water and sodium chloride were available ad libitum. Potassium concentrations were determined by atomic absorption spectroscopy. Plasma K concentration remained constant (3.49 +/- 0.09 mM K/L of plasma; mean +/- SEM) throughout the study. Erythrocyte potassium concentration decreased from 93.10 +/- 1.94 mM K/L of erythrocytes on day...
Three monoclonal antibodies identifying antigens on all equine T lymphocytes, and two mutually exclusive T-lymphocyte subsets. The aim of this study was to produce monoclonal antibodies (mAb) recognizing equine lymphocyte surface antigens. Fusions were conducted using BALB/c mice hyperimmunized with equine thymocytes. Hybridoma supernatants were screened by flow cytometry and positive hybridomas were cloned twice by limiting dilution. These mAb were then characterized for tissue distribution by immunohistology and flow cytometry, and by precipitation and analysis of the lymphocyte antigens which they recognized. Three mAb (CVS5, CVS4 and CVS8) are described which recognize only T lymphocytes in peripheral blood. Two-c...
Electron transfer between horse ferritin and ferrihaemoproteins. Reactions of reduced horse spleen ferritin with horse and Saccharomyces cerevisiae ferricytochromes c, cow ferricytochrome b5, sperm-whale metmyoglobin and Pseudomonas aeruginosa ferricytochrome c-551 were investigated by u.v.-visible spectrophotometry. In all cases the reduced ferritin reduced the ferrihaemoproteins. The rate of reduction varied from less than 0.2 M-1.s-1 for metmyoglobin to 1.1 x 10(3) M-1.s-1 for horse ferricytochrome c (0.1 M-phosphate buffer, pH 7.4, at 25 degrees C). We conclude that the mechanism of ferrihaemoprotein reduction involves long-range electron transfer throu...
Clinical and biochemical features of grass sickness (equine dysautonomia). An attempt has been made to assess the diagnostic value of clinical features seen at initial examination of horses with grass sickness, colic cases and cases submitted as possible grass sickness but diagnosed subsequently as some other condition. There appears to be no single pathognomonic sign for grass sickness. A range of signs has been associated with grass sickness but these are of value only when related to the length of illness and the history. Biochemical tests related to intestinal tissue damage, stress and dehydration were evaluated and most were found to be of value in diagnosing ac...
Equine cutaneous amyloidosis derived from an immunoglobulin lambda-light chain. Immunohistochemical, immunochemical and chemical results. Amyloid deposits from equine cutaneous nodular amyloidosis associated with extramedullary plasmacytoma were classified immunohistochemically as equine immunoglobulin lambda-light chain-derived and designated eA lambda (HIP). For chemical identification, the amyloid fibril proteins were separated on Sephadex G-100 in 6M guanidine.HCl. Polypeptides of predominantly 24 kDa and 50 kDa were found by polyacrylamide gel electrophoresis. They have preponderance of immunoglobulin lambda-antigenic determinants as detected by immunodiffusion and immunoblotting. Since the N-terminus of the major proteins ...
Clinical investigations of halothane and isoflurane for induction and maintenance of foal anesthesia. Fifty-eight foals were divided into two groups for study of aspects of the clinical anesthetic management of foals and to characterize effects of halothane (n = 30) and isoflurane (n = 28) in foals. There were no significant differences (P greater than 0.05) in the demographics of the two groups. Results of hemograms and biochemical analysis of venous blood samples before and after anesthesia were either not influenced or only mildly (clinically unimportant) affected by either agent. Like adult horses, foals have an increased PaCO2 when anesthetized with inhaled anesthetics. We could detect no...
Effects of chlorhexidine gluconate and chlorous acid-chlorine dioxide on equine fibroblasts and Staphylococcus aureus. Equine fibroblasts and Staphylococcus aureus were exposed for 30 minutes to six dilutions of chlorhexidine gluconate, a chlorous acid-chlorine dioxide irrigation solution, a chlorous acid-chlorine dioxide disinfectant, and phosphate buffered saline controls. Cell viability was determined by trypsinizing the cells, staining them with trypan blue, and counting cells that did not take the stain. All fibroblasts were killed when exposed to 1.0% and 0.5% chlorhexidine. The survival rate of fibroblasts increased linearly with decreasing concentrations of chlorhexidine gluconate, with a peak survival...
Ferritin: the role of aluminum in ferritin function. We previously showed that human brain ferritin (HBF) binds aluminum (Al) in vivo and in vitro and HBF isolated from Alzheimer's brain had more Al bound compared to aged matched controls (7). To further understand the role ferritin may play in Al neurotoxicity, we have studied in vitro the effect of Al on the function of human ferritin isolated from Alzheimer's (AD) and normal brain tissue, and compared the results with other mammalian ferritins. Al causes a concentration-dependent decrease in the initial rate of iron loading into apo-horse spleen and human brain ferritin and the rates were sim...
Oestrogen production by the preimplantation donkey conceptus compared with that of the horse and the effect of between-species embryo transfer. Aromatase distribution in membranes of preimplantation horse and donkey conceptuses was compared by measuring the incorporation of [3H]androstenedione into oestrone and oestradiol-17 beta. In the donkey conceptus, aromatase activity was similar in all the tissues examined (yolk sac, chorionic girdle and allantochorion), whereas in the horse it was generally lower and showed the relationship chorionic girdle greater than yolk sac greater than allantochorion. A higher proportion of labelled precursor was incorporated into oestradiol-17 beta by extra-embryonic tissues of the donkey compared with ...
Determination of short-chain fatty acids in equine caecal liquor by ion exchange high performance liquid chromatography after solid phase extraction. A high performance liquid chromatography (HPLC) method was developed for the determination of seven short-chain fatty acids in equine caecal liquor. Samples were cleaned up on a Sep-pak (C18) cartridge, and the analyte was eluted from the extraction cartridge and filtered through a 0.45 micron cellulose nitrate filter. The analyte was chromatographed by ion exchange HPLC. Detection was by UV at 210 nm. Recovery from phosphate buffer (0.05 M, pH 7.0) and equine caecal liquor was 76.95% (lactic), 76.76% (valeric). The limit of (propionic), 89.35% (isobutyric), 88.73% (butyric), 80.33% (isovaleri...
The isolation, characterisation and quantification of the equine plasma lipoproteins. Plasma lipoproteins were isolated from eight Thoroughbred horses and eight Shetland ponies on the basis of particle size by gel filtration chromatography and according to density using rate-zonal ultracentrifugation. Three major classes corresponding to very low density lipoproteins (VLDL), low density lipoproteins (LDL) and high density lipoproteins (HDL) were identified and characterised by their lipid and apolipoprotein compositions. The particle size distributions of each class were determined by electron microscopy and non-denaturing polyacrylamide gradient gel electrophoresis. HDL was fo...
