Proteins are essential macromolecules that play diverse roles in the physiology and health of horses. They are composed of amino acids and are involved in various biological processes, including tissue growth, repair, and the synthesis of enzymes and hormones. Dietary proteins are a key component of equine nutrition, influencing muscle development, immune function, and overall performance. Horses require a balanced intake of essential amino acids, which must be obtained through their diet, as they cannot be synthesized endogenously. This page compiles peer-reviewed research studies and scholarly articles that explore the types, functions, and dietary requirements of proteins in horses, as well as their impact on equine health and performance.
Cymbaluk NF, Christison GI, Leach DH.Eighteen Quarter Horse and Paint weanling colts were assigned to two treatments, either limit or ad libitum feeding. Nutrient digestibilities of diets were evaluated between 6 and 12 mo, 12 and 18 mo and 18 and 24 mo of age. From 6 to 12 mo, energy digestibility (64 to 68%) was unaffected by feed intake level. Ad libitum-fed horses had energy digestibilities 9 to 10% greater (P less than .05) than limit-fed horses between 12 and 24 mo of age, but adjustment for DM intake removed these differences. Limit-fed, mature ponies used dietary energy similarly to growing horses from 6 to 18 mo of age, ...
Hurd H.A total of 28 components were detected in the free amino acid (FAA) pool of hydatid fluid from primary and secondary equine cysts, secondary ovine cysts and host plasma. Examination of data from equine cysts revealed that the majority of FAAs were present in significantly greater concentrations in secondary cysts, glycine being over 30 times more concentrated. Values for total carbohydrates and glucose did not, however, differ significantly and total protein content was greater in primary cysts. Comparison of the (FAA) pool of secondary equine and ovine cysts revealed strain variation. It was ...
Lofstedt J, Jakowski RM.A 15-month-old Appaloosa colt had pyrexia, weight loss, diarrhea, and dependent edema. Abnormal laboratory findings included anemia, hypoalbuminemia, and hyperglobulinemia. Activities of liver-specific enzymes in this colt were high. Analysis of a liver biopsy specimen resulted in a presumptive diagnosis of tuberculosis. We recommend that liver biopsy be used as a diagnostic procedure if the differential diagnosis in a horse includes avian tuberculosis or granulomatous enteritis, especially if serum activities of liver-specific enzymes are high.
Saudek V, Atkinson RA, Williams RJ, Ramponi G.It has been proposed that combination of intraresidue, sequential and longer range nuclear Overhauser enhancements occurring in 1H nuclear magnetic resonance spectra of protein chains folded in a helix show a regular characteristic pattern. As a test case the spectra of horse muscle acylphosphatase were searched for this pattern together with other typical signs of a helical conformation (i.e. chemical shift, coupling constants and slow 2H-H exchange). Two amino acid sequences complying with these requirements were found. Just a few amino acid spin system assignments were then sufficient to lo...
Lombardo D, Chapus C, Bourne Y, Cambillau C.Horse (Equus caballus) pancreatic lipase (EC 3.1.1.3) has been crystallized using the hanging drop method of vapour diffusion at 20 degrees C. The best crystals were grown from an 8 mg/ml solution in 10 to 20% (w/v) polyethylene glycol 8000, 10 mM-MgCl2, 0.1 M-NaCl, 0.1 M-Mes buffer (pH 5.6). They reach dimensions of 0.8 mm x 0.4 mm x 0.6 mm. X-ray examination of the lipase crystals shows that they are orthorombic with a space group P2(1)2(1)2(1). Their cell dimensions are a = 79.8 A, b = 97.2 A c = 145.3 A. Two molecules per asymmetric unit give a Vm value of 2.82 A3/dalton (56% water content...
Saudek V, Williams RJ, Ramponi G.The solution structure of acylphosphatase determined by proton nuclear magnetic resonance spectroscopy is described. The results allow us to discuss the fold of the protein (101 amino acids), to correlate the exposure and the mobility of the backbone with the antigenicity, and to locate the active site.
Juneja RK, Sandberg K, Kuryl J, Gahne B.Two-dimensional agarose gel (pH 8.6)-horizontal polyacrylamide gel (pH 9.0) electrophoresis of horse serum samples, followed by general protein staining, revealed genetic polymorphism of an unidentified protein tentatively designated serum protein 3 (SP3). The SP3 fractions appeared distinctly when a 14% concentration of acrylamide was used in the separation gels. The 2-D mobilities of SP3 fractions were quite similar to that of albumin. Family data were consistent with the hypothesis that the observed SP3 phenotypes were controlled by four co-dominant, autosomal alleles (D, F, I, S). Evidence...
Patterson SD, Bell K.An affinity method was developed to investigate the interaction between protease and protease inhibitor by incorporating a protease incubation step into a two-dimensional electrophoretic separation of the plasma protease inhibitory proteins. This involved the application of the isoelectric focusing gel to filter paper saturated in the protease of choice before being placed on the second-dimensional polyacrylamide electrophoresis gel. General protein staining or immunoblotting was used to detect the protein or ligand in the complex. An in situ oxidation method was developed using the reagent ch...
Vaughn DM, Smyth GB.The serotonin metabolite, 5-hydroxyindoleacetic acid (5-HIAA) and the dopamine metabolite, homovanillic acid (HVA) in the cerebrospinal fluid (CSF) of seven clinically normal horses were evaluated with reverse phase high pressure liquid chromatography and electrochemical detection. Comparisons of the neurotransmitter metabolite concentrations were made on CSF collected simultaneously from the atlanto-occipital and lumbosacral regions. There were significantly higher amounts of 5-HIAA and HVA in atlanto-occipital CSF than in lumbosacral CSF. Mean 5-HIAA concentrations in atlanto-occipital and l...
dos Santos MC, D'Império Lima MR, Furtado GC, Colletto GM, Kipnis TL, Dias da Silva W.Pooled horse plasma containing antibodies against Crotalus durissus terrificus whole venom were digested with pepsin at an enzyme-substrate ratio of 8:1, pH 3.1, for 40 min and the F(ab')2M fragments purified by adding 8.7% caprylic acid (pH 5.0). For comparison, F(ab')2B purified by precipitation with ammonium sulphate and uncleaved IgG purified with caprylic acid were also prepared. Fab' fragments were obtained by reduction and alkylation of F(ab')2B. The anti-whole C.d. terrificus venom titers, determined by Dot-Blot were 12,800 (IgG), 6400 [F(ab')2B], 4800 [F(ab')2M] and 3200 (Fab'B). Immu...
Le Goff D, Pastier D, Hannan Y, Petit E, Ayrault-Jarrier M, Nouvelot A.1. Equine lipoproteins were isolated from plasma by density gradient ultracentrifugation and apolipoprotein composition determined by SDS-polyacrylamide gel electrophoresis. 2. VLDL and IDL were present at low concentration (0.2 mg/ml). Two apoB components of Mr corresponding to human apoB-100 and one apoB-48-like component were represented in VLDL fraction. 3. LDL-1 and LDL-2 subfractions have displayed an almost equal concentration (0.4 mg/ml). Two apoB-100-like components were the major apolipoproteins in each fraction. Small amounts of apoB-48-like component were detectable in LDL-1 and LD...
Juneja RK, Gahne B, Lukka M, Ehnholm C.By using immunoblotting with antiserum specific to human plasma apolipoprotein A-IV (apoA-IV), a previously reported polymorphic plasma protein of dogs viz postalbumin-2 (Pa2) and one of horses viz serum protein 2 (SP2), were identified as apoA-IV of these species. This along with earlier published results implied that: (1) both dog and horse show a high degree of polymorphism at the APOA4 locus with three common alleles in each of the two species; and (2) apoA-IV phenotyping in these two species can be done by analysing plasma/serum samples by a simple method of two-dimensional electrophoresi...
López ML, Grez P, Gribbel I, Bustos-Obregón E.The epididymis of stallion castrated during the breeding and non breeding seasons were subdivided into six regions and their ultrastructural and cytochemical characteristics were studied in order to provide a better understanding of the structure-function relationship of this androgen target organ. Even when the stallion has been postulated to be a seasonal breeder, our results do not show significant ultrastructural or cytochemical differences in both seasons. The pseudostratified epithelium is composed mainly of principal and basal cells and intraepithelial lymphocytes. The principal cells s...
Jolly RD, Dalefield RR.The lipopigments are a heterogenous group of pigments whose pathogenesis and terminology is confused. Whereas there is epidemiological and observational evidence that ceroid is derived from degeneration and peroxidation of unsaturated lipid, the assumption that all so-called lipopigments are similarly formed, is questioned. In particular, recent studies have distanced the pathogenesis of the pigment found in the ceroid-lipofuscinoses from that perceived for ceroid. The importance of protein rather than lipid in the pathogenesis of the pigment of ceroid-lipofuscinosis and of age pigment from th...
Hesford F, Lazary S, Curty-Hänni K, Gerber H.A number of horse alloantisera were characterized biochemically as being directed against MHC class I or class II antigens by immunoprecipitation of the corresponding antigens from lysates of biosynthetically radioactively labelled lymphocytes and determination of their molecular weights by SDS-PAGE and fluorography. Sera recognizing A2 and A3 specificities precipitated antigens of 44,000 Daltons molecular weight (class I heavy chain), whereas sera with specificities W13, W22 and W23 precipitated antigens corresponding to class II dimers (30,000 and 32,000 Daltons). Comparison with antigens pr...
Björkman C.1. Comparative data are presented of the activities of pyruvate dehydrogenase complex and acetyl-CoA synthetase and of the acetate content in homogenates from ventral grey matter in spinal cord from cows and two non-ruminant species, pigs and horses. The methods used in the study are evaluated and discussed. 2. The total pyruvate dehydrogenase complex activity was 24.9-29.9 mU/mg protein and did not differ between the species. The part of the complex that was in active form at the sampling occasion was 60, 85 and 95% in cows, pigs and horses, respectively. 3. Acetyl-CoA synthetase activity dif...
Izraely H, Choshniak I, Stevens CE, Demment MW, Shkolnik A.Factors determining the digestive efficiency of donkeys were studied in animals fed either a low quality roughage (wheat straw: 77.1% neutral detergent fibre, 2.8% crude protein) or a high quality forage (alfalfa hay: 47.5% neutral detergent fibre, 22.7% crude protein). The neutral detergent fibre (NDF) intake when fed wheat straw was 1693 +/- 268 g animal-1 day-1, 10% higher than when fed alfalfa hay. Digestive coefficient of NDF and acid detergent fibre (ADF) when fed wheat straw amounted to 50.9 +/- 4.9 and 42.0 +/- 4.1% respectively. NDF and ADF apparent digestibilities and mean retention ...
Jacob RJ, Steiner MR.Equine herpesvirus 3 (EHV-3)-infected equine cells display a kinetics of infected cell polypeptide (ICP) synthesis at 34 degrees C that is typical of coordinate cascade gene regulation of herpesviruses. In contrast, when infected cell cultures are incubated at the restricted temperature of 39 degrees C, the shift from early (beta) gene expression to late (gamma) gene expression is perturbed, i.e., there is an accumulation of early (beta) gene products and a decrease in, or absence of, late (gamma) gene products. Some of the affected late (gamma) gene products may be glycoproteins since these I...
Saad B, Corradin G, Bosshard HR.The antigenic determinants for two monoclonal antibodies directed against horse apo-cytochrome c, a protein of disordered structure, as judged by spectroscopic and hydrodynamic criteria, have been studied by a combination of methods: antigen competition in solution by radio immunoassay and enzyme-linked immunoassay, and differential acetylation of free and antibody-bound antigen. In the latter method the accessibility of lysine residues of the antigen in the antigen-antibody complex is compared to the accessibility in the free antigen. The two antibodies against the heme-free protein do not re...
Torres JL, Rush RS, Main AR.The serine carboxylesterase from horse serum was characterized by amino acid composition, peptide mapping, molecular and subunit weights, and sequencing of the amino acids around the essential serine residue at the active site. A protocol was developed for using reversed-phase high-performance liquid chromatography as the final step to obtain homogeneous preparations of horse serum carboxylesterase. Amounts sufficient for determining the amino acid composition and for peptide maps were obtained from a partially purified starting material which contained approximately 55% carboxylesterase. The ...
Lea RG, Bolton AE.Two-dimensional crossed immunoelectrophoresis of sera from pregnant and non-pregnant horses, using antisera developed against early pregnant mare serum, revealed the presence of two immunologically related proteins one of which appeared to be specific to the pregnant state. This pregnancy-specific protein had beta 2-electrophoretic mobility and was first detectable at Day 6 after successful mating with a stallion. The second protein had gamma 2-electrophoretic mobility and was present in sera from pregnant and non-pregnant horses. The proteins were termed beta 2-horse pregnancy protein and gam...
Mäenpää PE, Pirskanen A, Koskinen E.Serum alkaline phosphatase activity and serum concentrations of osteocalcin, free thyroxine, protein, and cholesterol were measured in 14 foals monthly from birth to 1 year of age. A transient decrease (P less than 0.05) in alkaline phosphatase activity and osteocalcin, free thyroxine, and cholesterol concentrations was found in November after transferring the horses from pasture to stables for the winter months. During the 2 subsequent months, serum alkaline phosphatase activity and osteocalcin concentration increased slightly. Protein concentration was highest in late fall and decreased (P l...
Soma LR, Behrend E, Rudy J, Sweeney RW.The disposition of flunixin meglumine administered IV at a dosage of 1.1 mg/kg was described by a 2-compartment model; the alpha and beta half-lives (t1/2) were 0.61 and 1.5 hours, respectively. When administered IV at a rate of 2.2 mg/kg, the disposition was best described by a 3-compartment model, and the alpha, beta, and lambda t1/2 were 0.16, 1.52, and 6.00 hours, respectively. The zero-time plasma concentrations after flunixin meglumine was administered at 1.1 and 2.2 mg/kg were 9.3 +/- 0.76 and 21.5 +/- 7.4 mg/L, respectively. The bioavailability after oral administration of 1.1 mg/kg wa...
Alvinerie M, Houin G, Toutain PL.The binding of prednisolone to total plasma proteins of dogs, horses, cows, and sheep was characterized using equilibrium dialysis. Prednisolone was bound to a first protein with high affinity but low capacity (transcortin) and to a second protein according to a nonsaturable mechanism (albumin). Interspecies differences were observed, with cows and dogs exhibiting the lowest, and sheep and horses the highest specific binding capacities. The results are in good agreement with known pharmacokinetic properties of prednisolone in domestic species.
Benanchi PL, Gazzei G, Giannozzi A.A simple two-step procedure for purifying F(ab)2 fragments of horse immunoglobulins is described. In the first step, the horse plasma is diluted, made up to 12% (w/v) with ammonium sulphate and digested with pepsin. In the second step, the previously dialyzed solution is chromatographed. Instead of a normal ion-exchange resin, a DEAE-cellulose, covalently linked to a synthetic vinyl polymer, was used (DEAE-Zeta-Prep). With this assembly it is possible to perform chromatography at a high flow-rate without the problems related to the use of large columns. The yield and purity of the final produc...
Godovac-Zimmermann J, Conti A, Napolitano L.The complete primary structure of donkey lysozyme has been established by pulsed liquid-phase sequencing of tryptic and chymotryptic peptides isolated by RP-HPLC. The positions of the Cys residues were identified by labeling the Cys residues with DABIA-reagent. Donkey lysozyme is a c-type lysozyme which is 129 amino acids long. It exhibits 50% homology to the human protein. We observe the full Ca(II) binding site suggested for the homologous alpha-lactalbumines. Although horse lysozyme has been reported to contain asparagine in position 61, which was in conflict with the three-dimensional stru...
Thompson KN, Jackson SG, Baker JP.Effects of high dietary levels of energy, protein and Ca and deficient levels of Ca on skeletal growth and development in the equine were studied in two experiments. The weanlings were fed grain-based diets. In Exp. 1, Group 1 (Ctl) received all nutrients at NRC recommended levels for growth; Group 2 (HE) received 150% of their digestible energy (DE) requirement; Group 3 (LC) received 150% of DE and 35% of Ca requirements. In Exp. 2, Group 1 (Ctl) again received all nutrients at NRC recommended levels for growth; Group 2 (HEP) received 150% of DE and 275% of CP requirements; Group 3 (HEPC) rec...
Robertson AT, Caughman GB, Gray WL, Baumann RP, Staczek J, O'Callaghan DJ.Equine herpesvirus type 1 (EHV-1) gene expression is coordinately regulated in an alpha, beta, gamma fashion. Viral alpha gene products include a 6.0-kb immediate early (IE) mRNA species (W. L. Gray et al., 1987, Virology 158, 79-87) and at least four closely related IE polypeptides (IEPs) (G.B. Caughman et al., 1985, Virology 145, 49-61). In this report, we describe results obtained from a series of in vitro translation experiments which were performed in an effort to characterize the IEPs and identify the mechanism by which individual IE protein species are generated. Our data indicate that ...
Allen D, Korthuis RJ, Clark ES.Microvascular permeability characteristics were evaluated in digits of 7 adult horses. After capillaries were isolated and an extracorporeal perfusion circuit for the digit was established, a lymphatic vessel draining the distal portion of the phalangeal region was cannulated at the level of the coronary band. Venous pressure was increased in a stepwise manner, and lymph flow, lymph protein concentration (Cl), and plasma protein concentration (Cp) were determined after measured variables were allowed to reach steady state. Lymph-to-plasma protein concentration ratios (Cl/Cp) and lymph and plas...
Miller PA, Lawrence LM.Six mature Quarter Horse mares were used in a crossover design to assess the effect of dietary protein level on metabolic response to exercise. After a 2-wk adaptation period to either a 12.9% (control) or an 18.5% CP (high-protein) diet, each mare performed a standard exercise test. The test consisted of a 15-min exercise period at 4.5 m/sec on a 9% grade motorized treadmill. Exercise resulted in an increase in heart rate (P less than .001), but there was no difference (P less than .05) between dietary treatments. In the jugular vein, lactate increased (P less than .01) from 6.3 to 52.0 mg/10...
van den Heuvel LP, van den Born J, Veerkamp JH, Janssen GH, van de Velden TJ, Monnens LA, Schröder CH, Berden JH.1. Proteoglycans extracted from human and equine glomerular basement membranes (GBM) were purified by ion-exchange chromatography and gel filtration. 2. The glycoconjugates had an apparent molecular mass of 200-400 kDa and consisted of 75% protein and 25% glycosaminoglycan. Glycosidase and HNO2 treatment and the amino sugar and sulfate composition of both proteoglycan preparations identified heparan sulfate (HS) as the predominant saccharide chain. 3. Hydrolysis with trifluoromethanesulfonic acid yielded comparable core proteins with molecular masses of ca 160 and 120 kDa. 4. The HS chains had...
Henry M, Amaral D, Tavares FF, Santos NR.The aim of the present study was to analyse the effect of hydrocoele of the vaginal cavity of stallions on semen characteristics, libido and fertility, and to contribute to the understanding of the aetiopathology of this condition in stallions. Hydrocoele did not substantially affect either the libido or semen characteristics of affected stallions compared with a control group, and had no effect on fertility. However, the protein content of hydrocoelic fluid was significantly higher than that of peritoneal fluid and was lower than the protein content of blood serum. Electrophoretic profiles of...
Fontanals AM, Becú T, Polledo G, Gaskin CK, Braun M.An R. equi vaccine, prepared under conditions which induce the expression of many antigens, and which has given encouraging results in field trials, was analyzed by SDS-PAGE and immunoblots and compared with other R. equi preparations: a preparation made in with the same technique from a nonvirulent isolate (virulence associated protein negative, VapA-negative); a whole cell preparation of a VapA-positive R. equi, prepared as a standard bacterin; and a semipurified VapA preparation (APTX). The antigens in these preparations were analyzed using hyperimmune sera (from adult horses vaccinated wit...
Matczuk AK, Chodaczek G, Ugorski M.Equine arteritis virus (EAV) is a prototype member of the Arterivirus family, comprising important pathogens of domestic animals. Minor glycoproteins of Arteriviruses are responsible for virus entry and cellular tropism. The experimental methods for studying minor Arterivirus proteins are limited because of the lack of antibodies and nested open reading frames (ORFs). In this study, we generated recombinant EAV with separated ORFs 3 and 4, and Gp3 carrying HA-tag (Gp3-HA). The recombinant viruses were stable on passaging and replicated in titers similar to the wild-type EAV. Gp3-HA was incorpo...
Kent JE.The concentrations of haptoglobin, immunoglobin (Ig)G(T) and IgG were measured in the serum of four previously parasite-free pony yearlings following a single dose of 700 (Group H) or 200 (Group L) stage three Strongylus vulgaris larvae (L3) and following a reinfection with the same doses 34 weeks later. The results are compared with an uninfected control pony. The haptoglobin concentration increased during Weeks 1 to 6 and 14 to 17 after infection in the serum of the ponies receiving 200 L3, but in only one pony dosed with 700 L3 (during Weeks 1 to 16). The serum haptoglobin also increased du...
Sato F, Yamashita S, Kugo T, Hasegawa T, Mitsui I, Kijima-Suda I.To determine the full-length complementary DNA (cDNA) sequence of equine erythropoietin (EPO) and to develop region-specific antibodies to differentiate equine EPO (eEPO) and human EPO (hEPO). Methods: RNA and lysate extracted from renal tissues of an adult Thoroughbred. Methods: Full-length cDNA was determined by use of a reverse transcriptase-polymerase chain reaction assay and a rapid amplification of cDNA ends method. The deduced amino acid sequence was compared with sequences of EPO reported for other species. Furthermore, 4 synthetic peptides were designed in 2 distinctive parts of the e...
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...
Miller L, Woodward EM, Campos JR, Squires EL, Troedsson M.The objectives of this study were to (i) verify localization of SP22 on fresh, cooled, and frozen/thawed equine spermatozoa and to (ii) determine SP22 mRNA and protein expression in equine testicular and epididymal tissues. Immunocytochemistry and Western blots were performed on the spermatozoa samples. Northern blots and Western blots were performed on the tissue samples. The immunocytochemistry revealed the presence of SP22 in all samples tested. The fresh spermatozoa stained predominantly over the equatorial segment as did the samples cooled for 1 and 2 days. The samples cooled for 3 days, ...
Desantis S, Albrizio M, Ventriglia G, Deflorio M, Guaricci AC, Minoia R, De Metrio G.The presence of the mu-opioid receptor and the type of glycosylation in the third extra-cellular loop of this receptor was investigated in the isthmus of mare oviduct during oestrus by means of immunoblotting and immunohistochemistry combined with enzymatic (N-glycosidase F and O-glycosidase) and chemical (beta-elimination) treatments. Immunoblotting analysis showed that the mu-opioid receptor consists of two peptides with molecular weights of around 65 and 50 kDa. After N-deglycosylation with N-glycosidase F an additional immunoreactive peptide was observed at around 30 KDa. The cleavage of O...
Tunón AM, Rodríguez-Martínez H, Hultén C, Nummijärvi A, Magnusson U.Undiluted uterine fluid from 20 Warmblood/Standardbred mares (5 to 14 yr old) was recovered by absorption to an intrauterine tampon. The mares were considered gynecologically healthy based on a clinical examination including uterine swabs for cytology and bacteriology as well as endometrial biopsy examinations. The protein profiles (SDS-PAGE) and concentrations of total protein, albumin, and immunoglobulins (Ig) A and G in the uterine fluid were examined and compared with the same proteins in serum. Major peaks were identified on the obtained protein profiles, and there was a clear similarity ...
Wasfi IA, Abdel Hadi AA, Elghazali M, Boni NS, Alkatheeri NA, Barezaig IM, Al Muharami AM, Hamid AM.The pharmacokinetics of tripelennamine (T) was compared in horses (n = 6) and camels (n = 5) following intravenous (i.v.) administration of a dose of 0.5 mg/kg body weight. Furthermore, the metabolism and urinary detection time was studied in camels. The data obtained (median and range in brackets) in camels and horses, respectively, were as follows: the terminal elimination half-lives were 2.39 (1.91-6.54) and 2.08 (1.31-5.65) h, total body clearances were 0.97 (0.82-1.42) and 0.84 (0.64-1.17)L/h/kg. The volumes of distribution at steady state were 2.87 (1.59-6.67) and 1.69 (1.18-3.50) L/kg, ...
Patterson SD, Bell K, Shaw DC.1. The donkey postalbumin protein has been shown to be the equivalent of human alpha 1 B-glycoprotein by protein immunoblotting and N-terminal amino acid sequence. 2. The horse A1B system (already identified as the homologue of human alpha 1 B-glycoprotein) and the donkey alpha 1 B-glycoprotein were characterized further for terminal sialic acid content, isoelectric point, amino acid composition and affinity for the dye-ligand, Cibacron Blue F3GA (known to bind human alpha 1 B-glycoprotein). 3. Two new alleles in the horse A1B system were found, bringing the total number of alleles to five. No...
Coté N, Trout DR, Hayes MA.Binding between equine plasma alpha-2-macroglobulin (alpha 2M) and several cytokines known to participate in inflammatory reactions in other species was initially examined. Plasma was obtained from 5 horses with various abnormalities. Samples, both untreated and after reaction with methylamine, were incubated with exogenous, radiolabeled, porcine-derived transforming growth factor-beta-1 (125I-TGF-beta 1), recombinant human interleukin-1-beta (125I-IL-1 beta), and recombinant human tumor necrosis factor-alpha (125I-rhTNF-alpha). They were then subjected to nondenaturing polyacrylamide gel elec...
Amann RP, Cristanelli MJ, Squires EL.Motility and fertility of frozen-thawed semen differs greatly amongst stallions. Differences in seminal plasma might be one cause of this variation. For 8 ejaculates from each of 17 stallions, seminal plasma was saved at -20 degrees C and spermatozoa were cryopreserved. Based on post-thaw sperm motility, seminal plasma samples from 7 stallions (2 good, 3 variable, 2 poor sperm motility) were selected for measurement of electrolytes, protein content and analysis by sodium dodecylsulphate gel electrophoresis (10% gel, Coomassie blue stain). Variation in seminal plasma was significant (P less tha...
Thomas PG, Ball BA, Ignotz GG, Dobrinski I, Parks JE, Currie WB.Before fertilization, equine spermatozoa adhere to oviduct epithelial cells (OEC) of the mare. The biochemical basis for this adhesion has not been determined. Our objective was to produce an antiserum to block this interaction. Ejaculated spermatozoa were subjected to nitrogen cavitation and spermatozoal plasma membranes enriched by sucrose density gradient centrifugation; membrane enrichment was confirmed by comparative alkaline phosphatase analysis, electron microscopy, and one- and two-dimensional PAGE. Periacrosomal plasma membrane was used as an immunogen for the production of an antiser...
Uberti B, Pressler BM, Alkabes SB, Chang CY, Moore GE, Lescun TB, Sojka JE.To investigate the effects of heparin administration on urine protein excretion during the developmental stages of experimentally induced laminitis in horses. Methods: 13 horses. Procedures-Horses received unfractionated heparin (80 U/kg, SC, q 8 h; n=7) or no treatment (control group; 6) beginning 3 days prior to induction of laminitis. All horses were given 3 oligofructose loading doses (1 g/kg each) at 24-hour intervals and a laminitis induction dose (10 g of oligofructose/kg) 24 hours following the final loading dose (designated as 0 hours) via nasogastric tube. Serum glucose and insulin c...
Weerasinghe CU, Learmonth GS, Gilkerson JR, Foote CE, Wellington JE, Whalley JM.The envelope glycoprotein D of EHV-1 (EHV-1 gD) is essential for virus infectivity and entry of virus into cells and is a potent inducer of virus-neutralizing antibody. In this study, truncated EHV-1 gD (gDt) was expressed with a C-terminal hexahistidine tag in E. coli using a pET vector. Western blot analysis using an anti-gD monoclonal antibody demonstrated the presence of gDt bands at 37.5, 36, 29.5 and 28 kDa. The immunogenicity and protective efficacy of partially purified gDt was compared with gD expressed in insect cells by a recombinant baculovirus (Bac gD) using a BALB/c mouse model o...
Urschel KL, Geor RJ, Hanigan MD, Harris PA.Stable isotope infusion methods have not been extensively used in horses to study protein metabolism. The objectives were to develop infusion and sampling methodologies for [1-(13)C] phenylalanine and apply these methods to determine whether the addition of supplemental amino acids to a control diet affected whole-body phenylalanine kinetics in mature horses. Arabian geldings were studied using a 6-h primed (9 μmol/kg), constant (6 μmol · kg(-1) · h(-1)) i.v. infusion of L-[1-(13)C] phenylalanine, with blood and breath sampled every 30 min, to measure whole-body phenylalanine kinet...
Shen Y, Ulaangerel T, Ren H, Davshilt T, Yi M, Li X, Xing J, Du M, Bai D, Dugarjav M, Bou G.The follicular fluid and oviduct fluid play major roles in oocyte maturation, sperm activation, and fertilization. To better understand the physiological environments for equine oocyte maturation and fertilization, here we conducted the proteome analysis and comparison on follicular fluids and oviduct fluids from the ovulatory side and the anovulatory side. The results showed that there is no significant difference between two side oviduct fluids, but a total of 71 differential abundance proteins (DAPs) were identified between two side follicular fluids, of which 9 are up-regulated and 62 are ...
Blanchard DB, Leid RW.Bovine and equine factor D were purified to apparent homogeneity as evidenced by a single protein staining band on 7.5-17.5% SDS-PAGE slab gels under both reducing and non-reducing conditions. An apparent mol. wt of 15,000 for bovine D and 22,500 for equine D were noted after SDS-PAGE gel analysis of both reduced and non-reduced preparations. A single polypeptide chain for both proteins was evidenced by the lack of any change in the electrophoretic mobility under each of these conditions. The bovine and equine D were enriched 3347- and 9447-fold, with a 20 and 29% yield of hemolytic activity, ...
Manso Filho HC, McKeever KH, Gordon ME, Costa HE, Watford M.One of the hallmarks of insulin resistance is a reduction in glucose transporter-4 (Glut-4) expression in adipose tissue but not in skeletal muscle. However, while Glut-4 has been demonstrated in skeletal and cardiac muscles in horses it has not been demonstrated in adipose tissue. The initial objectives of the present study were: (1) to test the hypothesis that Glut-4 expression would vary between selected key skeletal muscles; (2) to test the hypothesis that it would also vary between representative adipose tissue depots, and (3) to see whether expression would be greater in adipose tissue c...
Willis P, Sekhar KN, Brooks P, Fayrer-Hosken RA.To characterize further the events involved in fertilization and early embryonic development in the mare, effect of the estrous cycle on oviductal fluid proteins was investigated. Five mares had indwelling cannulas placed in their oviducts so that fluid could be collected throughout the estrous cycle. Daily fluid volumes were recorded and mares were monitored for signs of standing estrus. Oviductal fluid samples were pooled across mares according to stage of cycle (either estrus or nonestrus) for further analysis. Two-dimensional polyacrylamide gel electrophoresis (PAGE) was used to determine ...
Peixoto Rabelo I, Barroco de Paula V, Carvalho Bustamante C, Santana AM, Gomes da Silva D, Baldassi AC, Canola PA, Araújo Valadão CA.Horses submitted to carbohydrate overload can develop laminitis due to changes in cecal pH and microbiota, followed by an increase in transmural absorption of luminal content, including bacterial toxins. In response to acute injury there is hepatic overproduction of several proteins known as acute phase proteins (APP). Few studies have evaluated protein fractionation to characterize the inflammatory response in acute laminitis. The aim of this study was to test the viability of an experimental model to induce acute laminitis, using a single carbohydrate overload, and the influence of a bufferi...
Rathnam P, Fujiki Y, Landefeld TD, Saxena BB.Six hundred milligrams of follicle-stimulating hormone (FSH), containing 110 NIH-FSH-S1 units/mg, was isolated from 9 kg of equine pituitary glands. The equine FSH was dissociated into alpha- and beta-subunits. A tentative amino acid sequence of the alpha-subunit was determined. The alpha-subunit contained 82 amino acids. The equine FSH-alpha is shorter by 10 to 14 amino acids at the NH2 terminus and has several substitutions at several positions as compared with human FSH-alpha and bovine thyroid-stimulating hormone-alpha. At the intraspecies level, the alpha-subunits of human FSH, human lute...
Patterson SD, Auer D, Bell K.The induction of an acute phase response in four horses by adjuvant administration was used to examine the effect on the levels of plasma proteins. Blood parameters (packed cell volume, total plasma protein, red blood cell count, haemoglobin concentration) were monitored to follow the progress of the acute phase response in parallel with the examination of plasma proteins. Plasma protein levels were determined by densitometry from the electrophoretic patterns of three different gel systems. Haptoglobin and alpha 1 B glycoprotein were shown to be positive acute phase reactants whereas albumin w...
Semanchik PL, Wesolowski LT, Ryan PJ, White-Springer SH, Fluckey JD.Skeletal muscle is a highly dynamic organ that is essential for locomotion as well as endocrine regulation in all populations of horses. However, despite the importance of adequate muscle development and maintenance, the mechanisms underlying protein anabolism in horses on different diets, exercise programs, and at different life stages remain obscure. Mechanistic target of rapamycin (mTOR) is a key component of the protein synthesis pathway and is regulated by biological factors such as insulin and amino acid availability. Providing a diet ample in vital amino acids, such as leucine and gluta...
Saikku A, Koskinen E, Sandholm M.IgG levels and tryptic inhibition were investigated in sequentially collected mare's colostrum and milk, foal serum and urine. The colostral trypsin-inhibitor was "transfused" to the newborn foal by the colostral intestinal route in parallel with IgG. However, the trypsin-inhibitor as a small molecular weight inhibitor became excreted into urine peaking at about 20 hours. The physiological proteinuria in foals during the first 2 days is mostly due to immunoglobulin fragments and colostral-derived trypsin-inhibitor. Analysis of urine for IgG light chains or trypsin inhibitor will therefore reve...
DeBoer ML, Martinson KL, Kuhle KJ, Sheaffer CC, Hathaway MR.The impact of forage species on plasma amino acid (AA) concentrations of grazing horses (Equus caballus L.) is unknown. The objectives of this study were to determine the impact of different forage species on plasma AA concentrations and protein synthesis. Research was conducted in July in St. Paul, MN, USA. Alfalfa (Medicago sativa L.), mixed perennial cool-season grasses (CSGs), and teff (Eragrostis tef [Zucc.] Trotter) pastures were grazed by six horses randomly assigned to one of three forage types in a replicated Latin-square design. Horses had access to pasture each day. Jugular venous b...
Holterman WF, de Voogt P, Peereboom-Stegeman JH.Cadmium and zinc were determined in kidney cortex of 63 horses and 51 red deer (Cervus elaphus). Cadmium and zinc were also determined in protein fractions obtained by Sephadex chromatography of kidney cortex from 10 horses and 4 red deer. Histopathological parameters in kidney cortex of horses were compared to cadmium content. The metal contents (on wet weight basis) in kidney cortex of the horses were 0.31 +/- 0.22 mmole Cd/kg (range 0.03-1.21) and 0.63 +/- 0.17 mmole Zn/kg (range 0.36-1.23). The Zn content increased with the Cd content, the Zn increase being less at higher concentrations. N...
Hashimoto M, Nambo Y, Kondo T, Watanabe K, Orino K.In mammal circulation, ferritin-binding proteins (FBPs) are thought to be involved in clearance of circulating ferritin after complex formation with it through receptor-mediated uptake. However, there is no report on fetal FBP in fetal circulation. Although iron concentrations of fetal horse plasma were higher than those of adult horse plasma, plasma ferritin concentrations and ferritin-binding activities were found to be significantly lower in fetus than in adult. FBPs were purified from fetal or adult horse plasma on horse spleen ferritin-Sepharose 4B affinity column. Partially affinity-puri...
