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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.
[Histochemical and biochemical changes in skeletal muscles of rhabdomyolysis-sensitive racehorses following exertion. III: Elevated activity of various antioxidant enzymes]. In this communication, the results of a histochemical and biochemical enzyme study on gluteus medius muscle of horses, sensitive to exertional myopathy, during attacks of rhabdomyolysis are presented. For the biochemical study the biopsy specimens investigated were selected by means of histological and enzyme histochemical staining methods. Dissected specimens were used which contained groups of muscle fibres with a high or low activity of glucose-6-phosphate dehydrogenase. The activity of glucose-6-phosphate dehydrogenase, phosphogluconate dehydrogenase, glutathione reductase, glutathione per...
Degree of correspondence between contractile and oxidative capacities in horse muscle fibres: a histochemical study. Samples taken from the middle gluteal muscle of 95 untrained adult horses of different ages and sex were subjected to histochemical analysis using the myosin adenosine triphosphatase (m-ATPase) and nicotinamide adenine dinucleotide tetrazolium reductase (NADH-TR) staining techniques. Fibres were classified into types I, IIA and IIB according to m-ATPase activity after preincubation at pH 4.4. The percentage of FT (Fast-Twitch Glycolytic) fibres and the proportion of IIB fibres with "high" and "low" oxidative capacity were determined in serial sections stained for NADH-TR. Statistical analysis ...
The carbohydrate side chains of the major plasma serpins of horse and wallaby: analyses of enzymatic and chemically treated (including ‘Smith degradation’) protein blots by lectin binding. The carbohydrate side chains of the major plasma serpins of the horse and wallaby have been characterized by lectin analyses of protein blots from two-dimensional gels using the major human plasma serpin, alpha 1-protease inhibitor, as a control. Eight lectins were used in the characterization in conjunction with enzymatic deglycosylation of complex and high mannose side chains, chemical desialylation and defucosylation, and one round of 'Smith degradation', all being performed on the nitrocellulose blots. Assuming a standard complex side chain structure, the results of the 21 lectin/treatment...
The isoelectric focusing properties of serum alkaline phosphatase in disease and following prednisolone and phenylbutazone administration in the horse. This study was undertaken to ascertain if the isoelectric focusing pattern of serum alkaline phosphatase (AP) from sick horses with high activity is useful for determining its tissue origin. The effect of oral prednisolone and phenylbutazone therapy on this enzyme in healthy horses was also investigated. The sick horses were divided into three groups: hepatic, intestinal and miscellaneous. All sera had approximately thirteen bands of AP activity when focused on agarose gels with a pH gradient of 3.5 to 9.5. All the horses in the liver disease group had greater than 65% of enzyme activity in ba...
Histochemical and biochemical observations on milk-fat-globule membranes from several mammalian species. A specific secretory product of the lactating mammary gland are triglyceride fat globules which are enveloped by a very complex membrane, the milk fat globule membrane (MFGM). In different mammalian species (man, rhesus monkey, horse, goat, sheep, cow, grey seal, camel, alpaka) the glycoproteins of this membrane have been analyzed by gel electrophoresis, Western blotting and lectin histochemistry. A remarkable intra- and interspecific variability of these glycoproteins has been detected pointing to so far unknown physiological adaptions, which may play a role in the intestine of the new born. ...
Functional and morphological stasis during molecular evolution. The evolutionary distance between two sets of proteins was estimated using the techniques of Miyata and Yasunaga (1980) and Kimura (1980). Human beta 2-microglobulin was compared with the homologous murine molecule, while human and equine alpha-globin were similarly treated. It was found that a large amount of molecular evolution has occurred in beta 2-microglobulin since its divergence from the common ancestor of mice and humans. Kimura's estimate of evolutionary distance, K, is 0.353, while those of Miyata and Yasunaga are KS = 0.708 and KA = 0.171. The respective values for human and equine...
Equine postanaesthetic myositis: a possible role for free radical generation and membrane lipoperoxidation. A method for the evaluation of total plasma antihydroxyl and antiperferryl activity is described. This method was applied to horse plasma obtained during halothane anaesthesia. In horses suffering from postanaesthetic myositis, a significant decrease in the antiperferryl activity was observed during anaesthesia particularly when the muscular compression produced by the weight of the horse was released. In the affected muscles, strong oxidants could therefore be generated during the reperfusion of the ischaemic muscles and might initiate membrane lipid peroxidation. This phenomenon could possib...
Development of diurnal rhythm in some metabolic parameters in foals. 1. The development of diurnal rhythm activity of FDPA, AspAT and A1AT and in levels of cortisol, T3 and T4 was observed in the blood serum of six foals. 2. The studies began when a foal was 7 days old and were repeated every month until foals reached 1 year of age. Blood samples were taken every 4 hr for one day each month. 3. As a control group four barren mares were used, kept and examined in the same conditions. 4. In mature mares, diurnal rhythms in activity of A1AT (acrophase at 2200 hr), AspAt (2400 hr) and cortisol (0630 hr) but in T3 only in summer months (acrophase at 0100 hr) were ob...
High-performance liquid chromatography/tandem mass spectrometry: its use for the identification of stanozolol and its major metabolites in human and equine urine. A screening procedure for the anabolic steroid stanozolol in human and equine urine was developed based on enzymatic hydrolysis, liquid-liquid extraction and reversed-phase liquid chromatography combined on-line with tandem mass spectrometry. The column effluent was introduced into the atmospheric pressure ionization source of a triple-quadrupole mass spectrometer via a heated pneumatic nebulizer liquid chromatograph/mass spectrometer interface. Abundant protonated molecular ions were generated by corona discharge ionization. Confirmation of stanozolol and several of its hydroxylated and dihyd...
Rapid determination of methandrostenolone in equine urine by isotope dilution liquid chromatography-tandem mass spectrometry. Urine samples were spiked with [17-methyl-2H3]methandrostenolone as internal standard and extracted with a mixture of dichloromethane and cyclohexane. The organic phase was concentrated and injected onto a short octyl-silica column (30 mm x 4.6 mm I.D.) for separation of methandrostenolone and 17-epimethandrostenolone. The effluent from the column was connected to a Sciex TAGA 6000E triple quadrupole mass spectrometer equipped with an atmospheric pressure ion source for sampling of ions generated by a heated pneumatic nebulizer with corona discharge ionization. This ion source produced abundan...
Primary structure of horse serotransferrin glycans. Demonstration that heterogeneity is related to the number of glycans and to the presence of N-acetylneuraminic acid and N-acetyl-4-O-acetylneuraminic acid. Three serotransferrin variants Tf 2a, Tf 4b and Tf 5b were isolated in an homogeneous form from a preparation of homozygous horse serotransferrin Tf 0. On the basis of the results concerning molecular mass determination and the carbohydrate analysis, it is concluded that the serotransferrin variant Tf 2a contains only one glycan while variants Tf 4b and Tf 5b contain two glycans. The structure of all of the glycans has been established by combining methylation analysis, mass spectrometry and 400-MHz 1H-NMR spectroscopy. From the obtained results, it appears that the two glycans of Tf 5b varian...
Transbilayer movement of phosphatidylserine in nonhuman erythrocytes: evidence that the aminophospholipid transporter is a ubiquitous membrane protein. A 31-32-kDa integral membrane protein has been previously identified in erythrocytes as the protein most likely to be responsible for the transbilayer movement of phosphatidylserine (PS) [Connor & Schroit (1988) Biochemistry 27, 848-851]. Using similar techniques, we have identified analogous proteins of identical molecular weights in bovine, equine, ovine, porcine, canine, caprine, and rhesus red blood cells. Similar to human red blood cells, all of the mammalian cells were able to specifically transport an exogenously supplied fluorescent PS analogue from their outer-to-inner membrane le...
Serum biochemical indicators of liver function in neonatal foals. Serum biochemical indicators of liver function were determined in healthy, age-matched foals during the first 270 days of life. Values were compared with those of healthy adult horses and with those determined on the day of birth (less than 12 hours old). Serum alkaline phosphatase, gamma-glutamyl transferase, and L-iditol dehydrogenase activities were increased during the first 2 weeks of life. Serum cholesterol, triglyceride, and total and unconjugated bilirubin concentrations peaked during this same period. During the early neonatal period (less than 12 hours old), globulin concentrations (...
Serum uric acid concentrations in horses heterozygous for combined immunodeficiency. Serum uric acid concentrations were determined in horses known to be carriers of combined immunodeficiency gene(s) and in presumed noncarrier horses. Uric acid concentrations were significantly higher (P less than 0.005) in carrier horses than in presumed noncarrier horses. However, there was some overlap in serum uric acid concentrations between carrier and presumed noncarrier horses.
Biotransformation of 5(10)-estrene-3 alpha,17 beta-diol by equine testicular preparations in vitro. The research investigates the biosynthesis of a particular isomer called 5(10)-estrene-3 alpha, 17 beta-diol in stallion testes and how it affects the formation of 19-nor steroids and oestrogens. Summary of […]
Effect of calcium-channel blockers and salbutamol on the isolated mare uterus–interaction with the calcium agonist Bay K 8644. The effects of nifedipine, verapamil and diltiazem were investigated in the isolated mare uterus in comparison with salbutamol. All the calcium-channel blockers and salbutamol inhibited the spontaneous, KC1- and electrically induced contractions; nifedipine and salbutamol were the most potent compounds. The calcium agonist Bay K 8644 (10(-8)-10(-6) mol/l) competitively antagonized the inhibitory effect of nifedipine (pA2 value = 8.54 +/- 0.06), whereas it was only slightly or totally ineffective against verapamil, diltiazem and salbutamol. These results indicate that calcium-channel blockers a...
In vitro transport of L-alanine by equine cecal mucosa. When sheets of mucosa from the cecum of clinically normal horses were incubated in vitro with radiolabeled L-alanine, they could accumulate this amino acid against an apparent concentration gradient after 60 to 150 minutes of incubation. The active transport system for L-alanine was on the serosal surface of the mucosal sheet only. L-Alanine accumulation at 60 minutes was partly inhibited by 20 mM glycine (P less than 0.01), 0.5 mM ouabain (P less than 0.05), and Na deprivation (P less than 0.02). Anoxia for 60 minutes increased L-alanine accumulation, but had adverse effects on cell structure...
Concentration and degree of polymerization of hyaluronate in equine synovial fluid. In addition to its well-known physicochemical properties, hyaluronate (HA) has recently been shown to have important biological and pathophysiologic regulatory effects on granulocytes, monocytes, fibroblasts, and endothelial cells, as well as on the healing of wounds and various joint disorders. Many of these effects depend on or are reflected in the concentration and degree of polymerization of HA. Therefore, high-performance liquid chromatography with size-exclusion column was used to characterize the concentration and degree of polymerization of HA in equine synovial fluid (SF). The mean (+...
Thin-layer chromatographic screening procedure for some drugs in horse plasma. A thin-layer chromatographic screening procedure for some basic, neutral and acidic drugs was developed using 3 ml of horse plasma. Chloroform-2-propanol (95:5, v/v) was used as the extraction solvent. The drugs were identified by a high-performance thin-layer chromatographic plate and spraying successively with some detection reagents. In this study, the extraction recovery rates and the detection limits were determined at the same time.
Simultaneous analysis of furosemide and bumetanide in horse plasma using high performance liquid chromatography. A high performance liquid chromatographic method is described for the simultaneous determination of furosemide and bumetanide in horse plasma. The C8 (3 microns) reversed phase column (4.8 x 150 mm) provided clear separation of furosemide and bumetanide with other components present in the horse plasma. The detection limit for both the drugs was 10 ng/mL. Both drugs were stable in plasma (at natural or acidic pH) for up to 24 h. The method is sufficiently sensitive to detect furosemide levels in plasma obtained from horses receiving a therapeutic dose of furosemide.
An improved TLC method for the detection of flunixin in equine serum. A method for flunixin detection in equine serum extracts involving thin layer chromatography, spraying the chromatogram with alkaline sodium hypochlorite solution and heating with a detection limit of 50 ng ml-1 is described.
Histochemical and molecular determination of fiber types in chemically skinned single equine skeletal muscle fibers. Until now, there has been no reliable method for histochemical determination of fiber types of single skinned muscle fibers. The major problem arises from the fact that most histochemical techniques use cross-sections of a large group of fibers and compare a given fiber with those surrounding it. This is not possible with a single skinned fiber which has been separated from a bundle to be used for mechanical analysis. A further problem is that the skinning procedure itself may alter the staining pattern. We have developed a procedure by which multiple cross-sections of single skinned fibers ca...
Functional and morphological studies on blood platelets in a thrombasthenic horse. A four-year-old Standardbred gelding presented with a 3.5 year history of intermittent epistaxis and spontaneous submucosal petechiae and ecchymoses in the nares and the mouth. Routine haematological and biochemical examinations were unremarkable. A thrombocytopathy was suspected when activated partial thromboplastin time, one stage prothrombin time, plasma fibrinogen and the platelet count were all normal. The patient's platelets failed to aggregate with serotonin, adenosine diphosphate, collagen (at 20 micrograms/ml) or the endoperoxide analogue U46619. Very high levels of collagen (100 micr...
Altered sarcoplasmic reticulum function after high-intensity exercise. This study examined the effects of acute high-intensity exercise on the rate and capacity of Ca2+ uptake and Ca2+-stimulated adenosinetriphosphatase (ATPase) activity of the sarcoplasmic reticulum and the reversibility of these effects. Thoroughbred horses were run at maximal O2 uptake on a high-speed treadmill until fatigued. Muscle temperatures and biopsy samples were collected at rest, immediately after exercise, and 30 and 60 min after exercise. Blood samples were collected at rest and 5 min after exercise. Muscle and blood (lactate concentration) were three- and fivefold greater than pre-...
Androgen and 19-norandrogen aromatization by equine and human placental microsomes. The ability of equine and human placental microsomes to aromatize testosterone and 19-nortestosterone was studied. When 3 microM [1 beta,2 beta-3H]testosterone was used as substrate, the specific activity of equine placental microsomal aromatase was 2.5 times higher than that of the human microsomal enzyme. Although 19-nortestosterone was aromatized 67 times more rapidly by equine than by human aromatase, we found that equine aromatase exhibited a markedly weaker affinity for this substrate than did the human enzyme. Competitive inhibition of testosterone aromatization by 19-nortestosterone oc...
A new method for hydrolyzing sulfate and glucuronyl conjugates of steroids. A new method for hydrolyzing steroid conjugates (both sulfates and glucuronides conjugates) that is efficient, effective, and inexpensive is described. This method comprises incubation of the conjugates--after salting-out into ethyl acetate or elution from a C18 cartridge--with anhydrous methanolic hydrogen chloride (methanolysis) for 10 min. It has been successfully applied to our routine radioimmunoassay screening and GC/MS confirmation studies of steroids in prerace and postrace equine urine samples. Comparative GC/MS studies on entire (male horse) urine samples showed that methanolysis gav...
Comparison of the binding of Ca2+ and Mn2+ to bovine alpha-lactalbumin and equine lysozyme. The enthalpy change of the binding of Ca2+ and Mn2+ to equine lysozyme was measured at 25 degrees C and pH 7.5 by batch microcalorimetry: delta H degrees Ca2+ = -76 +/- 5 kJ mol-1, delta H degrees Mn2+ = -21 +/- 10 kJ mol-1. Binding constants, log KCa2+ = 6.5 +/- 0.2 and log KMn2+ = 4.1 +/- 0.5, were calculated from the calorimetric data. Therefore, delta S degrees Ca2+ = -131 +/- 20 JK-1 mol-1 and delta S degrees Mn2+ = 8 +/- 44 JK-1 mol-1. Removal of Ca2+ induces small but significant changes in the circular dichroism spectrum, indicating the existence of a partially unfolded apo-conformatio...
