Enzymes are biological catalysts that facilitate biochemical reactions in horses by lowering the activation energy required for these processes. They are involved in various physiological functions, including digestion, metabolism, and cellular repair. Common enzymes in equine biology include amylase, lipase, and lactate dehydrogenase, each playing a specific role in the breakdown of nutrients and energy production. The activity and concentration of these enzymes can vary in response to different physiological and pathological conditions, serving as potential indicators in veterinary diagnostics. This page compiles peer-reviewed research studies and scholarly articles that explore the function, regulation, and clinical implications of enzymes in equine health.
Gross MK, Toscano DG, Toscano WA.Calmodulin (CaM), the calcium binding protein that modulates the activity of a number of key regulatory enzymes, is present at high levels in sperm. To determine whether CaM regulates adenylate cyclase in mammalian sperm, the actions of EGTA and selected CaM antagonists on a solubilized adenylate cyclase from mature equine sperm were examined. The activity of equine sperm adenylate cyclase was inhibited by EGTA in a concentration-dependent manner with a half-maximal inhibitory concentration (IC50) of 2 mM. Equine sperm adenylate cyclase was also inhibited in a concentration-dependent manner by...
Shipton WA.The equine phycomycotic agent known commonly as Hyphomyces destruens or occasionally as Pythium gracile, is described as a new species Pythium destruens. Separation is on the basis of morphological features, temperature growth profiles, esterase/lipase activity, and carbohydrate utilization ability. P. diclinum (synonymous with P. gracile sensu Middleton) showed minor differences in vesicle, oospore and oogonium size from P. destruens. P. destruens grew at 40 degrees C on corn meal agar and hydrolysed esters of lauric and oleic acids. These abilities were not displayed by P. diclinum, but this...
Gaillard JL, Silberzahn P.In the stallion testis, aromatase activity was localized in the microsomal fraction. Androgen aromatization occurred through the loss of 1 beta,2 beta hydrogen atoms and appeared to involve free sulfhydryl groups. A single enzyme system seemed to aromatize androgen and norandrogen at the same rate while having a much lower affinity for norandrogens.
Yancik SA, McIlwraith CW, Wagner AE, Trotter GW.Creatine kinase (CK) and lactate dehydrogenase (LD) enzyme activities and isoenzymes were determined for synovial fluid, synovial membrane, and articular cartilage from 24 clinically normal equine tarsocrural (tibiotarsal) and femoropatellar joints. All 3 tissues contained LD isoenzymes LD1 to LD5, and CK isoenzymes BB and MM. The CK isoenzyme MB was not found. The similarities in isoenzyme composition of these 3 tissues made differentiation of the source of LD and CK impossible by isoenzyme pattern alone. Reference values for the total enzyme activities of specific joint tissues also had wide...
Berti A, Degl'Innocenti D, Stefani M, Liguri G, Ramponi G.A non competitive enzyme-linked immunosorbent assay (ELISA) specific for horse muscle acylphosphatase (E.C. 3.6.1.7.) has been developed. The purified anti-acylphosphatase antibodies were immobilized by passive absorption to a solid-phase support and incubated with known and unknown amounts of antigen. The antibody-acylphosphatase complex was quantified using the same antibody conjugated to horseradish peroxidase. The assay yields positive reactions with as little as 0.05 ng of antigen, with intra- and interassay coefficients of variation of 5% and 7%, respectively. On the basis of this assay ...
Sanny CG, Weiner H.The inhibition of mitochondrial (pI 5) horse liver aldehyde dehydrogenase by disulfiram (tetraethylthiuram disulphide) was investigated to determine if the drug was an active-site-directed inhibitor. Stoichiometry of inhibition was determined by using an analogue, [35S]tetramethylthiuram disulphide. A 50% loss of the dehydrogenase activity was observed when only one site per tetrameric enzyme was modified, and complete inactivation was not obtained even after seven sites per tetramer were modified. Modification of only two sites accounted for a loss of 75% of the initial catalytic activity. Th...
Scandurra R, Politi L, Santoro L, Consalvi V.Horse liver phosphopantothenoylcysteine decarboxylase (EC 4.1.1.36) incorporates nonexchangeable tritium from borotritide with a decrease of the activity. Substrate prevents both tritium incorporation and the decrease in activity. Acid and base hydrolysis of the tritiated protein releases labeled lactate identified by high-voltage paper electrophoresis, paper chromatography and silicic acid chromatography. These results indicate the presence of pyruvate covalently bound through an ester linkage to phosphopantothenoylcysteine decarboxylase which is then another example of a mammalian enzyme in ...
Archibald AL.1. Serum isoamylases with alpha-glucosidase activity from cattle, sheep, horses, goats, red deer, pigs and dogs were compared to one another. 2. The isoamylases from cattle and pigs were polymorphic. 3. In agarose gel electrophoresis the isoamylases behaved as alpha-1-globulins but in starch gel electrophoresis they were differentially retarded by affinity effects. 4. Molecular weights were estimated: cattle (417,000); sheep (402,000); horses (420,000); goat (399,000); red deer (405,000); pigs (375,000) and dogs (390,000). 5. Isoelectric points were estimated: cattle, sheep, goat and red deer ...
Andersson L, Arnason T, Sandberg K.Investigations on relationships between biochemical polymorphism and variation in quantitative traits are of interest from the perspectives of both theoretical quantitative genetics and practical animal breeding. This subject was studied by using racing performance records of more than 25,000 horses of the Swedish Trotter breed born in the period 1970-1979. For all horses data on six blood group and nine electrophoretic loci were available. Two different performance traits were investigated. A racing performance index value was calculated for all individuals which had started in at least five ...
Malmheden Yman I, Sandberg K.Meat from the species horse, donkey and their hybrids, mule/hinny, can be reliably identified by determination of genetic variants of serum albumin by starch gel electrophoresis of meat extracts. Staining of the starch gel for carboxylesterase activity permits differentiation of most horses from donkeys while mules/hinnies cannot be distinguished from horses by their esterase activity alone.
von Fellenberg R.The importance of proteases and protease inhibitors for the pathogenesis of pulmonary emphysema and chronic bronchitis of the horse is described. Endogenous elastases from neutrophil granulocytes and macrophages, which probably provoke emphysema in the human being, are not relevant in horse emphysema. Exogenous elastases from different species of streptomyces may be responsible for emphysema generation in this species. Part of the exogenous elastases are poorly or not inhibited at all by the equine blood protease inhibitors especially by alpha 1-protease inhibitors. A disorder similar to genet...
Clyne MJ.Proteoglycan degradation is central to the development of degenerative joint disease. Proteoglycans may be degraded by lysosomal enzymes from chondrocytes, synoviocytes or leucocytes. Collagen and matrix degradation occurs either by direct damage or due to degrading enzymes released into synovial fluid. Once the pathological sequence has begun it continues in a cyclic manner unless arrested by the ability of chondrocytes to synthesise sufficient matrix components. Treatment should ideally be directed to this end.
Numazawa M, Osawa Y.The metabolic pathway leading to equilin and equilenin biosynthesis in the pregnant mare is different from that of estrone and estradiol and it is apparently cholesterol-independent. The precise precursors and intermediates and the stereomechanism of equine placental aromatization have not been established. [1,2-3H, 4-14C]3-Hydroxy-3,5,7-androstatrien-17-one was synthesized as a potential substrate and the 3H-distribution was analyzed by biochemical and chemical derivatization methods. The substrate was converted to equilin, equilenin and Heard's ketone by horse placental microsomes with a sp....
Miller SK.1. Results of laboratory experiments which compared horse and human serum butyrylcholinesterase (EC 3.1.1.8) with respect to their acid inactivation and ammonium sulfate protection show: 2. Horse serum butyrylcholinesterase is more resistant to inactivation at pH 3.0 than human serum butyrylcholinesterase. 3. The loss of activity at pH 3.0 for both horse and human butyrylcholinesterase does not follow first order kinetics. 4. Both human and horse serum butyrylcholinesterase are protected from pH 3.0 inactivation by ammonium sulfate concentrations up to 33% saturation (1.37 M).
Pellegrini A, Hägeli G, von Fellenberg R.1. Trypsin digestion of perchloric acid precipitated horse plasma yielded polypeptides with inhibitory properties for trypsin, chymotrypsin and, to a small extent, kallikrein. 2. The Mr of the inhibitory polypeptides were 73,000 and 24,000. 3. The number, enzyme specificity and Mr of the inhibitory polypeptides differed from the values known for the human being. 4. The inhibitory polypeptides were purified by affinity chromatography on Sepharose-trypsin and by gel filtration through Sephadex G-75. 5. Protease inhibitory polypeptides were generated in the same manner by chymotrypsin, elastase, ...
Yamamoto M, Yamamoto I, Tanaka Y, Sugano M.1. A method for the purification of horse serum lecithin:cholesterol acyltransferase has been established. 2. The method involves the adsorption of the enzyme from diluted horse serum on DEAE-Sephadex A-50, (NH4)2SO4 fractionation, 1-butanol treatment, and chromatographic techniques of DEAE-Sepharose CL-6B, DEAE-Sephadex A-50, Affi-Gel blue and hydroxylapatite. 3. The resultant enzyme preparation essentially formed a single main band when subjected to polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. 4. The final purification of the enzyme was 20,000-fold with 7% yi...
Medda R, Cara N, Floris G.Ceruloplasmin is a blue copper-containing serum glycoprotein with oxidase activity. It as been proposed that the physiological function of ceruloplasmin involves the oxidation of ferrous iron and its incorporation into apotransferrin. There are several reports demonstrating that ceruloplasmin is made up of multiple chains. Ryden has questioned the multichain structure of ceruloplasmin from human, pig, horse and rabbit sera, arguing that the dissociation observed by previous workers could be attributed to cleavage of labile bands in the protein by enzymatic contaminants present in commercial pr...
Patterson WH, Brown CM.Serial blood samples were obtained from 16 Standardbred foals from time of birth to postpartum day 28. Sera were obtained and analyzed for gamma-glutamyltransferase (GGT), aspartate transaminase, and immunoglobulin (Ig) G. Presuckle colostrum from the respective mares of these foals was analyzed for GGT activity. Mean serum aspartate transaminase activities were significantly increased above presuckle values by postpartum hour 48 (P less than 0.01) and increased gradually over the first 14 days. Mean serum IgG concentrations were significantly greater than presuckle values by 5 hours after foa...
Frauenfelder HC, Rossdale PD, Ricketts SW, Allen WR.Blood samples were collected twice weekly over a nine month period from 24 Thoroughbred racehorses in training at Newmarket to study the effects of daily training schedules and stage of oestrous cycle on serum enzyme levels and clinical signs of equine exertional myopathy. The sampling period extended from November to July and was performed between 16:00 h and 18:00 h so as to be at least 6 h after exercise. Serum creatine kinase, aspartate aminotransferase and plasma progestogen concentrations were measured. All horses were subjected to a similar traditional training programme. The study demo...
Pomelova VG, Gaĭdamovich SIa, Demenev VA, Kadoshnikov IuP.A three-step concentration of Venezuelan equine encephalomyelitis (VEE) virus from tissue culture fluid was carried out in a two-phase system of polyethyleneglycol (PEG)--sodium dextran sulphate (SDS). The concentration method was based on the dependence of virus distribution coefficient upon NaCl content in the system which allowed alternating transfer of the virus from one phase of the system into the other. The infectious activity of the virus increased approximately 100-fold after the first step, 190-fold after the second, and 300-fold after the third step. The process of concentration was...
Poerio E, Parr GR, Taniuchi H.The previous studies (Juillerat, M. A., and Taniuchi, H. (1986) J. Biol. Chem. 261, 2697-2711), using a three-fragment complex (1-25)H X (28-38) X (39-104) of horse cytochrome c, have shown that invariant leucine 32 and partially invariant leucine 35, both buried in the interior, exhibit a striking difference in perturbation of binding fragment (28-38) by substitution with isoleucine. Then the idea has been proposed that the energy states of leucine 32, the Met-80-S-heme-Fe bond and other distant residues such as tryptophan 59 would be coupled to generate extra force while leucine 35 would be ...
Andrews FM, Spurgeon TL.The histologic and histochemical staining characteristics of the triceps brachii (long head), extensor carpi radialis, gluteus medius, vastus lateralis, biceps femoris, semimembranosus, semitendinosus, and extensor digitorum longus muscles of 8 Thoroughbreds, 2 Quarter Horses, 1 Arabian, 1 Paso Fino, and 1 Shetland Pony are described. Muscle fiber morphology, staining distribution and intensity, amount of IM connective tissue, number of IM blood vessels and IM nerves, calcium-activated adenosine triphosphatase activity (CaATPase), percentage of fibertype population, percentage of relative fibe...
Lessard P, Wilson WD, Olander HJ, Rogers QR, Mendel VE.Twenty horses of various ages had inadvertently ingested alfalfa hay contaminated with Senecio vulgaris. Among them, 4 died of liver disease. Blood was collected from affected horses at monthly intervals for 7 months and at the 9th and 14th months. The following serum enzymes and chemical items were assayed: aspartate aminotransferase, lactate dehydrogenase, alkaline phosphatase, gamma-glutamyl transferase, sorbitol dehydrogenase, total bilirubin, BUN, glucose, cholesterol, inorganic phosphate, calcium, total protein, and albumin. Amino acid profiles, conjugated bile acids, sulfobromophthalein...
Bayly WM, Brobst DF, Elfers RS, Reed SM.Serum and urinary biochemical changes were recorded in 5 ponies in which acute tubular nephrosis had been induced over 5 days with mercuric chloride and potassium dichromate. Serum osmolality, the serum concentrations of urea nitrogen, creatinine, sodium, potassium and chloride, and blood pH and blood gases were measured daily for 14 days or until humane euthanasia was performed. Levels of the same substances were quantitated daily in urine. In addition, routine urinalyses and determination of urinary gamma glutamyl transferase (GGT) activity were performed on each sample. Changes in the value...
Brobst DF, Carroll RJ, Bayly WM.Urinary concentrations of gamma glutamyl transferase (GGT), alkaline phosphatase (AP), aspartate aminotransferase (AsAT), and alanine aminotransferase (AAT) were measured in 32 healthy horses (16 geldings and 16 females) at the same time on 2 consecutive days. The subjects were divided into 4 ages groups, each comprising 8 horses (4 of each sex). In 10 of the geldings, urine was collected continuously for 72 hours, with catheterized samples being obtained at 0800, 1400 and 2000 hours, and an aliquot of the pooled urine being taken every 24 hours. Urinary enzyme activity was found to be unrelat...
Bauer E, McDougall J, Cameron BD.Isomerization of trans-4'-(2-hydroxy-3,5-dibromo-benzylamino)cyclohexanol (HDBC) in vivo has been investigated in horse, cow, dog, rat and man. Following oral administration of 4'-trans-HDBC to the horse, a very efficient first-pass trans-cis isomerization was observed. In the urine of the horse and cow, 40% and 29% respectively of the conjugated alcohols consisted of the 4'-cis isomer. Isomerization in rat and dog took place only to a small extent, and in man no 4'-cis isomer was detected. Oxidation of HDBC to the corresponding ketone, at pH 9.0, was highest with horse- and rat-liver 10 000 g...
Weers JG, Maki AH.Triplet-singlet energy transfer has been studied in the complex formed between auramine O (AO) and horse liver alcohol dehydrogenase with optically detected magnetic resonance (ODMR) spectroscopy. The results show that Trp-15 and Tyr residues transfer triplet energy mainly by a trivial process, whereas Trp-314 transfers triplet energy by a Förster process with two observed lifetimes at 77 K of 170 and 50 ms. The different Förster energy-transfer lifetimes are ascribed either to quenching of the two Trp-314 residues of the dimer by a single asymmetrically bound AO or to two distinct conformat...
Strambini GB, Gonnelli M.The inactivation of equine liver alcohol dehydrogenase by guanidine hydrochloride and urea has been studied by monitoring the intrinsic tryptophan fluorescence and phosphorescence emission. The use of triplet-state lifetimes to probe the flexibility of protein structure at the site of tryptophan-314 reveals a distinct behavior between the two denaturants. At predenaturational concentrations, the loss of enzyme activity in guanidine hydrochloride is associated with a loosening of intramolecular interactions resulting in a greater fluidity of the interior region of the macromolecule. In contrast...
Lamar CH, Turek JJ, Bottoms GD, Fessler JF.Certain in vitro culture conditions were determined for equine endothelial cells obtained from the aorta and pulmonary arteries. Cells were enzymatically isolated from the vessel lumen, using clostridial collagenase (2.5 mg/ml of Hanks's balanced salt solution) incubated at 37 C for 30 minutes. Cells were cultured in alpha minimum essential medium supplemented with plasma-derived and nonplasma-derived bovine fetal sera, endothelial cell-growth supplement, heparin, and antibiotics. Smooth muscle cell growth was not inhibited with nonplasma-derived animal sera, plasma-derived equine serum, or he...
Art T, Votion D, Lekeux P.The purpose of this study was 1) to measure and compare some physiological parameters in horses during a standardised treadmill exercise performed either in temperate atmospheric conditions (TC) (ambient temperature: 15 degrees C; relative humidity: 55%), or in hot and humid conditions (HHC) (ambient temperature: 30 degrees C; relative humidity: 75%) and 2) to follow the recovery of the same horses during 1 h after both tests. Five healthy fit Standardbred horses were investigated twice at 8 days interval, in TC and in HHC. Some measurements were made during a standardised treadmill exercise t...
de Kok PM, Beijer NA, Buck HM, Sluyterman LA, Meijer EM.The geometry of seven NAD+ analogues bound to horse liver alcohol dehydrogenase (LADH) modified only in their nicotinamide group, have been studied using AMBER molecular mechanics energy-minimization procedures. Starting geometries were taken from X-ray crystallographic data for NAD+/Me2SO/LADH reported by Eklund and co-workers. In this study the NAD+ analogues were encaged by the constituent amino acids of the enzyme within a range of 0.6 nm from the initial NAD+/Me2SO/Zn2+ complex. The calculational method used is able to rationalize individual substituent effects and to evaluate the essenti...
Kareskoski AM, Reilas T, Sankari S, Andersson M, Güvenc K, Katila T.Seminal plasma (SP) is a mixture of contents from the testes, epididymides and accessory sex glands. The sperm concentration is highest in the first few jets, or fractions, of the ejaculate, and the composition of SP varies between these fractions because accessory gland secretions are released in a specific order. The aim of this study was to compare the levels of Na, Cl, K, Mg, Ca, inorganic phosphate (Pi) and the enzymes alkaline phosphatase (AP), acid phosphatase (ACP) and β-glucuronidase (BG) in the different fractions of the ejaculate and in different stallions. All semen collections we...
von Fellenberg R.The importance of proteases and protease inhibitors for the pathogenesis of pulmonary emphysema and chronic bronchitis of the horse is described. Endogenous elastases from neutrophil granulocytes and macrophages, which probably provoke emphysema in the human being, are not relevant in horse emphysema. Exogenous elastases from different species of streptomyces may be responsible for emphysema generation in this species. Part of the exogenous elastases are poorly or not inhibited at all by the equine blood protease inhibitors especially by alpha 1-protease inhibitors. A disorder similar to genet...
Brobst DF, Carroll RJ, Bayly WM.Urinary concentrations of gamma glutamyl transferase (GGT), alkaline phosphatase (AP), aspartate aminotransferase (AsAT), and alanine aminotransferase (AAT) were measured in 32 healthy horses (16 geldings and 16 females) at the same time on 2 consecutive days. The subjects were divided into 4 ages groups, each comprising 8 horses (4 of each sex). In 10 of the geldings, urine was collected continuously for 72 hours, with catheterized samples being obtained at 0800, 1400 and 2000 hours, and an aliquot of the pooled urine being taken every 24 hours. Urinary enzyme activity was found to be unrelat...
West HJ.In 18 horses there was no effect of age or sex on plasma activities of gamma-glutamyl transferase (gamma-GT), 5'-nucleotidase (5'-NT) and leucine aminopeptidase (LAP). All the enzymes were equally stable after storage for one month at -20 degrees C and there was no significant difference between their activities in serum and plasma in clinically normal horses. The pattern of release of gamma-GT, 5'-NT and LAP into plasma was studied in 114 horses which had a variety of orthopaedic, gastrointestinal, cardiovascular and hepatic (necrosis, lipidosis, neoplasia and cirrhosis) conditions. A definit...
Hoffman M, Cheong SH, Stokol T.Liver analyte measurement is important in the evaluation of sick animals. Liver injury in horses is recognized by increased glutamate dehydrogenase (GLDH), sorbitol dehydrogenase (SDH), and aspartate aminotransferase (AST) activities, whereas biliary pathology is identified by increased alkaline phosphatase and γ-glutamyl transferase (GGT) activities or bilirubin concentrations. We have observed high GLDH, but not SDH, activities in neonatal foals admitted for conditions other than liver disease. Only one previous study have evaluated GLDH activity over time in healthy neonatal foals; however...
Schmitz A, Portier CJ, Thormann W, Theurillat R, Mevissen M.Stereoselectivity has to be considered for pharmacodynamic and pharmacokinetic features of ketamine. Stereoselective biotransformation of ketamine was investigated in equine microsomes in vitro. Concentration curves were constructed over time, and enzyme activity was determined for different substrate concentrations using equine liver and lung microsomes. The concentrations of R/S-ketamine and R/S-norketamine were determined by enantioselective capillary electrophoresis. A two-phase model based on Hill kinetics was used to analyze the biotransformation of R/S-ketamine into R/S-norketamine and,...
Marshall DE, Dumasia MC, Wooding P, Gower DB, Houghton E.Maternal endometrial and fetal allantochorionic tissues were separated manually from the placentae of seven healthy thoroughbred and three pony mares, ranging in gestational age from 100 to 318 days. The homogeneity of subcellular fractions prepared from these tissues was assessed initially using the marker enzymes, succinate dehydrogenase, NADPH cytochrome C reductase and lactate dehydrogenase for the mitochondrial, microsomal and cytosolic fractions, respectively. Light microscopy and histochemical analysis demonstrated that the separated fetal allantochorionic membrane, which is made up of ...
Barton MH, Darden JE, Clifton S, Vandenplas M.To validate primer sets for use in reverse transcription quantitative PCR assays to measure gene expression of cytosolic phospholipase A2 (cPLA2) and microsomal prostaglandin E2 synthase 1 (mPGES1) in equine mononuclear cells and determine the effects of firocoxib, a selective cyclooxygenase 2 (COX-2) inhibitor, on COX-2, cPLA2, and mPGES1 gene expression following incubation of mononuclear cells with lipopolysaccharide (LPS). Methods: 8 healthy adult horses. Methods: Peripheral blood mononuclear cells were isolated by density gradient centrifugation and incubated at 37°C with medium alone, f...
Storgaard IH, Kristensen J, Larsen C, Mertz N, Østergaard J, Larsen SW.Intra-articular depot injectables based on in situ suspension formation of ester prodrugs of nonsteroidal anti-inflammatory drugs are promising for management of joint pain. As candidates for this delivery approach, 5 diclofenac ester prodrugs comprising different imidazole-containing promoieties were synthesized and their physicochemical properties characterized. In vitro hydrolysis rates were investigated in buffer solutions, in 40% (v/v) human, equine, canine, and rat plasma, and in 80% (v/v) human and equine synovial fluid. Bioconversion of the prodrugs to diclofenac was found to be enzym...
Shellow JS, Jackson SG, Baker JP, Cantor AH.Twenty mature geldings, averaging 535 kg, were used to determine the influence of dietary selenium (Se) on the blood levels of Se and Se-dependent glutathione peroxidase (SeGSH-Px) activity in the horse. Horses were randomly assigned within breed to four treatments consisting of five horses each and fed a basal diet containing .06 ppm of naturally occurring Se. Diets were supplemented with .05, .10 and .20 ppm Se, as sodium selenite. Blood was drawn for 2 wk before, and for 12 wk following, the inclusion of supplement Se in the diets. Whole blood and plasma Se concentrations and plasma SeGSH-P...
Diel de Amorim M, Klein C, Foster R, Dong L, Lopez-Rodriguez MF, Card C.Leucyl and cystinyl aminopeptidase (LNPEP/oxytocinase) is an enzyme that metabolizes oxytocin in serum and tissues. The presence of oxytocin/neurophysin I (OXT), oxytocin and LNPEP and their relationship to other genes is unknown in the equine conceptus. Our objective was to characterize gene expression of LNPEP and OXT on D8, 10, 12, 14, 15, 16 and 21 conceptuses in relationship to other genes. Immunohistochemistry, western blot and liquid chromatography with tandem mass spectrometry (LC-MS/MS) were used for identification of oxytocin and LNPEP in D15, 16 and 18 conceptuses. LNPEP was increas...
Toscano WA, Toscano JS, Toscano DG, Gross MK.The soluble calmodulin-sensitive isoform of adenylyl cyclase isolated from equine sperm is unique because it requires Mn(2+) rather than Mg(2+) for activity. To gain insight into the molecular action of metals on sperm adenylyl cyclase, the kinetics of Mn(2+) and ATP effect was examined. A biphasic response to increases in ATP concentration was observed when metal was held constant. When [Mn(2+)] exceeded [ATP], however, greatly enhanced enzyme activity was observed. The kinetic profiles were consistent with allosteric activation of adenylyl cyclase by Mn(2+). Linear transformation of the data...
Kareskoski M, Vakkamäki J, Laukkanen K, Palviainen M, Johannisson A, Katila T.Matrix metalloproteinase (MMP)-2 and MMP-9 are gelatinases that take part in several reproductive processes. The aim of this study was to measure levels of MMP-2 and MMP-9 in fractionated stallion ejaculates, and to evaluate the association between these components and semen quality, and sperm longevity during cooled storage. Semen quality were assessed separately for sperm-rich fractions (HIGH), sperm-poor fractions (LOW), and whole ejaculate samples (WE) from 33 stallions. After cooled storage with SP either present or removed, sperm motility and DFI were determined. The relative activity of...
Horney BS, Honor DJ, MacKenzie A, Burton S.Serum sorbitol dehydrogenase (SDH) activities in 10 cows and nine horses were measured using an automated clinical analyzer. The serum samples were divided into aliquots that were stored at room temperature (21 degrees C), refrigerated (0-5 degrees C), or frozen (-30 degrees C). The stability of the SDH activity was monitored at various intervals. SDH activity in bovine sera remained stable for at least 5 hours at room temperature, 24 hours refrigerated, and 72 hours frozen without any significant (p < 0.05) differences from the initial serum values. In equine sera, SDH activity remained st...
Weers JG, Maki AH.Triplet-singlet energy transfer has been studied in the complex formed between auramine O (AO) and horse liver alcohol dehydrogenase with optically detected magnetic resonance (ODMR) spectroscopy. The results show that Trp-15 and Tyr residues transfer triplet energy mainly by a trivial process, whereas Trp-314 transfers triplet energy by a Förster process with two observed lifetimes at 77 K of 170 and 50 ms. The different Förster energy-transfer lifetimes are ascribed either to quenching of the two Trp-314 residues of the dimer by a single asymmetrically bound AO or to two distinct conformat...
Wasfi IA, Zorob OM, Boni NS, Hadi AA, Agha BA, Elghazali M.The activity of hepatic mixed function oxidases was compared in horses and camels (Camelus dromedarius) by studying the pharmacokinetics of antipyrine in seven camels and five horses following intravenous administration of a single dose of antipyrine (25 mg/kg). The data obtained (mean +/- SEM and median in brackets) in camels and horses, respectively, were as follows: the elimination half-lives were 3.25 +/- 0.23 (3.19) and 3.09 +/- 0.25 (2.90) hr; the apparent volumes of distribution (area method) were 0.691 +/- 0.045 (0.648) and 0.642 +/- 0.034 (0.676) l/kg; the volumes of distribution at s...
Desmadril M, Mitraki A, Betton JM, Yon JM.The unfolding-folding transition of phosphoglycerate kinase induced by GuHC1 was studied at equilibrium. Various signals were used to follow the transition: fluorescence emission, difference spectra, circular dichroism and enzymatic activity. The non-coincidence of transition curves obtained from different structural parameters indicate a deviation from a two-state process. The view that structural domains behave as independent "folding units" is critically discussed.
Rueca F, Conti MB, Porciello F, Spaterna A, Antognoni MT, Mangili V, Fruganti G, Avellini G.The purpose of this study was to assess the possible relationship between maximal running speed, serum isoenzyme patterns of creatine kinase (CK) and lactate dehydrogenase (LDH) and echocardiographic indices of left ventricular function. A group of 15 healthy, 3-year-old Maremmano stallions were given a 100 day training programme. At the end of this the animals carried out a maximum speed test and were divided into 2 groups (A and B) according to whether or not they had attained a speed of 15 m/s. Venous blood samples were taken from each horse before exercise (T0), 2 min (T1) and 24 h (T2) af...
Clemmons RM, Lee MR, Bliss EL, Asbury AC, Cook D, Brown V.Superoxide dismutase (SOD), a free radical scavenger with anti-inflammatory activity, was administered IM to horses. Ex vivo platelet aggregation in response to arachidonic acid was monitored to determine whether exogenous SOD altered equine platelet prostaglandin metabolism. Preparations of platelet-rich plasma obtained before SOD administration were incubated with different concentrations of SOD and were aggregated with arachidonic acid. Superoxide dismutase did not exert a demonstrable effect, either ex vivo or in vitro. Aspirin abolished arachidonic acid-induced platelet aggregation in vit...
Patterson SD, Bell K, Manton VJ.A detailed biochemical characterization of four of the five previously described alleles of the plasma protease inhibitor (Pi) system of Equus przewalskii was performed using both one- and two-dimensional electrophoretic techniques. The proteins have been characterized in terms of isoelectric point, relative molecular mass, inhibitory activity to bovine trypsin and chymotrypsin, immunochemical cross-reactivity, terminal sialic acid content and enzyme:inhibitor complex formation and the oxidation sensitivity of this interaction. Using these functional criteria, only three loci (Spi 1, 2 and 3) ...
Gaillard JL, Silberzahn P.In the stallion testis, aromatase activity was localized in the microsomal fraction. Androgen aromatization occurred through the loss of 1 beta,2 beta hydrogen atoms and appeared to involve free sulfhydryl groups. A single enzyme system seemed to aromatize androgen and norandrogen at the same rate while having a much lower affinity for norandrogens.
Sasaki K, Igarashi S, Amasaki T, Amasaki H, Nishita T, Kano Y, Asari M.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 ...
Afonso T, Giguère S, Rapoport G, Berghaus LJ, Barton MH, Coleman AE.Angiotensin-converting enzyme (ACE) inhibitors are used in horses with cardiovascular disorders despite the paucity of available data regarding their efficacy. Objective: The degree of serum ACE inhibition varies considerably between drugs. Methods: Eight healthy adult horses. Methods: Randomized prospective study. Horses were fasted overnight prior to receiving one of 4 ACE inhibitors intragastrically, administered at one of 2 dosages, using a randomized Latin square design (benazepril: 0.5 and 0.25 mg/kg; ramipril: 0.3 and 0.1 mg/kg; quinapril: 0.25 and 0.125 mg/kg; perindopril: 0.1 and 0.05...
BMC research notesNovember 8, 2012
Volume 5 626 doi: 10.1186/1756-0500-5-626
Boudko SP, Ishikawa Y, Lerch TF, Nix J, Chapman MS, Bächinger HP.Hyperelastosis cutis is an inherited autosomal recessive connective tissue disorder. Affected horses are characterized by hyperextensible skin, scarring, and severe lesions along the back. The disorder is caused by a mutation in cyclophilin B. Results: The crystal structures of both wild-type and mutated (Gly6->Arg) horse cyclophilin B are presented. The mutation neither affects the overall fold of the enzyme nor impairs the catalytic site structure. Instead, it locally rearranges the flexible N-terminal end of the polypeptide chain and also makes it more rigid. Conclusions: Interactions of th...
Gondim FJ, Modolo LV, Campos GE, Salgado I.Mammalian skeletal muscle expresses splice variants of neuronal nitric oxide synthase (nNOS). Skeletal muscles have a metabolically heterogeneous population of myofibers, and fiber composition in equine skeletal muscle is correlated with athletic ability in endurance events. In this study, we investigated whether nNOS expression in equine skeletal muscle is related to fiber type and endurance training. Biopsy samples obtained from the gluteus medius of sedentary- (SH) and endurance-trained (TH) horses were examined for the electrophoretic mobility of myosin heavy chain (MHC) and NOS activity. ...
O'Donnell MM, Harris HM, O'Toole PW, Ross RP.We report the draft genome sequence of Lactobacillus equi strain DPC6820, isolated from equine feces. L. equi is a predominant Lactobacillus species in the horse hindgut microbiota. An examination of the genome identified genes and enzymes highlighting L. equi adaptations to the herbivorous gastrointestinal tract of the horse, including fructan hydrolases. This genome sequence may help us further understand the microbial ecology of the equine hindgut and the influence lactobacilli have on it.
Wauters J, Pille F, Martens A, Franck T, Serteyn D, Gasthuys F, Meyer E.Equine joint infection is a life-threatening disorder, and confirmation of the diagnosis can be difficult. Synovial fluid biomarkers may assist the discrimination between infectious and noninfectious joint disease. Objective: This study investigates whether the immunological detection of total and enzymatically active myeloperoxidase (MPO) assists the diagnosis of joint infection in horses. Methods: The following 4 sample groups were included: healthy; osteochondritis dissecans (OCD); traumatic synovitis; and culture-confirmed infected joints. Synovial fluid was analysed for total MPO by a hor...
Wurster U, Riese K, Hoffmann K.An attempt was made to establish normal values for the total protein concentrations and the enzyme activities of LDH, MDH and PGI in the intraocular fluids of rats, guinea pigs, rabbits, cats, dogs, sheep, cattle, pigs, horses and humans. Remarkably little species differences were noted in 9 of the 10 mammals with vitreal enzyme activities falling into a narrow range between 8.4 U/l (PGI, horse) and 92.4 U/l (MDH, guinea pig). All species obeyed the sequence aqueous less than vitreous less than serum with exception of the rat, where vitreous activities surpassed serum at least two-fold. The ve...
