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.
Kang J, Lee MS, Watowich SJ, Gorenstein DG.A phosphorothioate RNA aptamer (thioaptamer) targeting the capsid protein of Venezuelan equine encephalitis virus (VEEV) was isolated by in vitro combinatorial selection. The selected thioaptamer had a strong binding affinity (approximately 7nM) and high specificity for the target protein. For the binding to the protein, the overall tertiary structure of the thioaptamer is required. We introduce two theoretical methods to examine the effect of phosphorothioate modification on the enhancement of binding affinity and one experimental method to examine the nature of the multiple bands of thioapta...
Boscan P, Watson Z, Steffey EP.To investigate the changes in colloid osmotic pressure (COP) and total protein concentrations during routine general anesthesia in horses. Methods: Prospective, clinical study. Methods: Twelve adult healthy horses aged 9.1 +/- 4.7 years and weighing 474 +/- 79 kg presented for elective surgery and 14 adult horses aged 8.7 +/- 7.3 years and weighing 510 +/- 85 kg. Methods: All horses were premedicated with xylazine and anesthesia induced with ketamine, diazepam and guaifenesin, and maintained with isoflurane for 2.5 hours. Lactate Ringer's solution was administered at 11 mL kg(-1) hour(-1). Osm...
Floyd RV, Mason SL, Proudman CJ, German AJ, Marples D, Mobasheri A.Aquaporins (AQPs) play fundamental roles in water and osmolyte homeostasis by facilitating water and small solute movement across plasma membranes of epithelial, endothelial, and other tissues. AQP proteins are abundantly expressed in the mammalian kidney, where they have been shown to play essential roles in fluid balance and urine concentration. Thus far, the majority of studies on renal AQPs have been carried out in laboratory rodents and sheep; no data have been published on the expression of AQPs in kidneys of equines or other large mammals. The aim of this comparative study was to determ...
Okamura M, Yokoyama N, Takabatake N, Okubo K, Ikehara Y, Igarashi I.In the present study, we investigated the effects of protease pretreatments of host erythrocytes (RBC) on the in vitro growth of bovine Babesia parasites (Babesia bovis and B. bigemina) and equine Babesia parasites (B. equi and B. caballi). The selected proteases, trypsin and chymotrypsin, clearly modified several membrane proteins of both bovine and equine RBC, as demonstrated by SDS-PAGE analysis; however, the protease treatments also modified the sialic acid content exclusively in bovine RBC, as demonstrated by lectin blot analysis. An in vitro growth assay using the protease-treated RBC sh...
McKeever KH, Agans JM, Geiser S, Lorimer PJ, Maylin GA.Recombinant human erythropoietin (rhuEPO) causes an increase in red blood cell production and aerobic capacity in other species; however, data are lacking on effects in the horse. Objective: This study tested the hypothesis that rhuEPO administration would alter red cell volume (RCV), aerobic capacity (VO2max) and indices of anaerobic power. Methods: Eight healthy, unfit mares accustomed to the laboratory and experimental protocols were randomly assigned to either a control (CON, n = 4; 3 ml saline 3 times/week for 3 weeks) or EPO group (EPO, n = 4, 50 iu/kg bwt rhuEPO/3 ml saline 3 times/week...
Eizema K, van der Wal DE, van den Burg MM, Dingboom EG, Everts ME.An optimal developed musculoskeletal system is vital for the performance of the horse. Previously, we showed that in the m. gluteus medius from adult untrained horses, identical mRNA and protein expression patterns were found in the majority of fibres. However, co-expression of IIa and IId/x myosin heavy chain (MyHC) was substantially more common at the protein than at the mRNA level, suggesting a transcriptionally controlled fine-tuning of these 2 genes. Objective: To analyse the MyHC transcripts and proteins (including the cardiac alpha isoform) in the same muscle during post natal developme...
Matsui A, Ohmura H, Asai Y, Takahashi T, Hiraga A, Okamura K, Tokimura H, Sugino T, Obitsu T, Taniguchi K.In man, muscle protein synthesis is accelerated by administering amino acids (AA) and glucose (Glu), because increased availability of amino acids and increased insulin secretion, is known to have a protein anabolic effect. However, in the horse, the effect on muscle hypertrophy of such nutrition management following exercise is unknown. Objective: To determine the effect of AA and Glu administration following exercise on muscle protein turnover in horses. We hypothesise that administration of AA and Glu after exercise effects muscle hypertrophy in horses, as already shown in man and other ani...
Stewart-Hunt L, Geor RJ, McCutcheon LJ.Increased insulin sensitivity occurs after a period of exercise training, but the mechanisms underlying this training-associated increase in insulin action have not been investigated. Objective: To examine the effects of short-term endurance training (7 consecutive days) and a subsequent period of inactivity (5 days) on whole body insulin sensitivity and GLUT-4 protein and the activities of glycogen synthase (GS) and hexokinase (HK) in skeletal muscle. It was hypothesised that training would increase insulin sensitivity in association with increased GLUT-4 protein and activities of GS and HK, ...
Connysson M, Muhonen S, Lindberg JE, Essén-Gustavsson B, Nyman G, Nostell K, Jansson A.High-energy forage might be an alternative to concentrates for performance horses and such forage can be produced by an early cut. However, early cut forage is high in crude protein (CP), which may result in an excessive CP intake. Objective: To investigate how CP intake affects nitrogen (N), fluid and acid-base balance, and exercise response in horses fed high-energy forage diets. The hypothesis was that high CP intake causes acidosis, and alters fluid balance and response to intensive exercise. Methods: Two forage-only diets based on high-energy grass forage were fed for 23 days in a crossov...
McCutcheon LJ, Geor RJ, Pratt SE, Martineau E, Ho K.Exercise is an important stimulus for the regulation of numerous metabolic functions in muscle, but there has been little investigation of the mechanism of exercise-enhanced glucose utilisation. Objective: To examine the effects of exercise on the signalling components Akt and glycogen synthase kinase-3 (GSK3) involved in insulin regulation of glycogen synthase (GS) activity. It was hypothesised that hyperinsulinaemia and prior exercise would alter the extent of Akt phosphorylation and GSK3beta deactivation and increase insulin stimulation of GS activity in skeletal muscle. Methods: Eight Stan...
Belogortseva N, Rubio M, Terrell W, Miksovská J.Photoacoustic calorimetry and transient absorption spectroscopy were used to study conformational dynamics associated with CO photodissociation from horse heart myoglobin (Mb) reconstituted with either Fe protoporphyrin IX dimethylester (FePPDME), Fe octaethylporphyrin (FeOEP), or with native Fe protoporphyrin IX (FePPIX). The volume and enthalpy changes associated with the Fe-CO bond dissociation and formation of a transient deoxyMb intermediate for the reconstituted Mbs were found to be similar to those determined for native Mb (DeltaV1 = -2.5+/-0.6 ml mol(-1) and DeltaH1 = 8.1+/-3.0 kcal mo...
Morte MI, Rodrigues AM, Soares D, Rodrigues AS, Gamboa S, Ramalho-Santos J.The goal of this work was to correlate oxidative stress caused by reactive oxygen species (ROS) and DNA damage with classic semen parameters in spermatozoa and seminal plasma of fertile and subfertile stallions. Oxidation was measured in both lipids and proteins, using the thiobarbituric acid reactive species (TBARS) assay and the DNPH carbonyl groups assay, respectively. Sperm DNA damage was monitored using the TUNEL assay. These parameters were monitored in samples obtained during the breeding and the non-breeding seasons. In general, fertile stallions showed better classical semen parameter...
Nic An Ultaigh S, Ryan MF.Proteinases released during the in vitro maintenance of asynchronous cultures of the free-living nematode Caenorhabditis elegans were characterized on the basis of subunit composition, fluorogenic substrate specificity, inhibitor sensitivity and pH optima. Cysteine proteinases are present in the excretory-secretory products (ESP) as indicated by the hydrolysis of cathepsin fluorogenic substrates and confirmed by immunoblotting. Serine proteinases were predominant as indicated by substrate gel analysis and inhibitor studies. The presence of metallo-proteinases was also indicated by inhibitor st...
Hejmej A, Kotula-Balak M, Sadowska J, Bilińska B.Connexin 43 (Cx43) is a ubiquitously distributed gap junction protein in testes and other reproductive tissues. Adjacent cells share ions and small metabolites through intercellular channels, which are present in gap junctions. Previously, Cx43 has not been reported in testes, epididymides and prostates either in healthy stallions or cryptorchid horses. Objective: To demonstrate the expression pattern of Cx43 in the reproductive tissues of stallions and examine whether naturally occurring bilateral cryptorchidism has any influence on distribution and expression of Cx43. Methods: The expression...
Guo HY, Pang K, Zhang XY, Zhao L, Chen SW, Dong ML, Ren FZ.This study investigated the changes in chemical composition, nitrogen fraction distribution, and AA profile of milk samples obtained during lactation from the Jiangyue breed of donkey in Northwest China. Results showed that donkey milk contained 9.53% total solids, 1.57% protein, 1.16% fat, 6.33% lactose, and 0.4% ash on average, which is more similar to mare and human milk than to the milk of other mammals. Throughout the lactation investigated, pH and density were constant, protein and ash content showed an apparent negative trend (an increase in lactose content during 120 d postpartum, foll...
Miclo L, Girardet JM, Egito AS, Mollé D, Martin P, Gaillard JL.Highly phosphorylated casein with a low molecular mass was isolated from Haflinger mare's milk by RP-HPLC. It accounts for 4.0% of the casein content. Its mass was determined by LC-ESI-MS before and after treatment by alkaline phosphatase. The molecular mass found for the apo-form (10,591 +/- 2 Da) is in agreement with its primary structure, which was established by ESI-MS/MS from tryptic peptides. It appeared that this short protein (94 amino acid residues) is an internally truncated form of the full-length equine beta-casein (226 residues). This low-Mr variant of equine beta-casein displays ...
Tiyawisutsri R, Holden MT, Tumapa S, Rengpipat S, Clarke SR, Foster SJ, Nierman WC, Day NP, Peacock SJ.The bacterial biothreat agents Burkholderia mallei and Burkholderia pseudomallei are the cause of glanders and melioidosis, respectively. Genomic and epidemiological studies have shown that B. mallei is a recently emerged, host restricted clone of B. pseudomallei. Results: Using bacteriophage-mediated immunoscreening we identified genes expressed in vivo during experimental equine glanders infection. A family of immunodominant antigens were identified that share protein domain architectures with hemagglutinins and invasins. These have been designated Burkholderia Hep_Hag autotransporter (BuHA)...
Timoney JF, Qin A, Muthupalani S, Artiushin S.Streptococcus equi, a clonal descendent of an ancestral S. zooepidemicus, causes equine strangles, a highly contagious purulent lymphadenitis of the head and neck. The aim of this study was to evaluate as vaccine components novel surface exposed or secreted S. equi proteins identified in an expression gene library with sera from resistant horses. Six proteins expressed by S. equi CF32 but not by S. zooepidemicus 631 were used to vaccinate one group of eight ponies. A second pony group was immunized with five adhesin and other proteins encoded by genes of Linkage Gr 1. All ponies made strong se...
Seeliger F, Hess O, Pröpsting MJ, Naim HY, Kleinschmidt S, Woehrmann T, Germann PG, Baumgärtner W.A 20-year-old female horse showed a nodular, firm, focal ulcerated mast cell tumor at the right dorsobuccal face of the tongue. Histologically, the nonencapsulated tumor consisted of dense, infiltrating aggregates of well-differentiated, Cresyl violet-positive mast cells accompanied by numerous eosinophils. Furthermore, they exhibited a strong, diffuse, intracytoplasmatic immunohistochemical signal for tryptase and a faint membrane-associated and perinuclear signal for tyrosine kinase receptor KIT. Confocal laser scanning microscopy confirmed an aberrant spatial colocalization of KIT in the Go...
Groot MT, Merkx M, Koper MT.The redox properties of horse and yeast cytochrome c electrostatically immobilized on carboxylic acid-terminated self-assembled monolayers (SAMs) have been determined over a broad pH range (pH 3.5-8) in the absence and presence of nitric oxide. Below pH 6, both proteins exhibit comparable midpoint potentials, coverages, and electron-transfer rate constants, which suggests that they are adsorbed on the SAM in a similar fashion. Above pH 6, a sharp decrease in electron-transfer rate constants is observed for immobilized yeast cytochrome c, which is indicative of a change in the electron tunnelin...
Li H, Zou X, Springer M, Briest A, Lind M, Bünger C.Randomized and self-controlled study with anterior lumbar interbody fusion in a porcine model. Objective: To determine the osteoinductive potential of an equine bone protein extract in anterior interbody spinal fusion. Background: Interbody spinal fusion with bone graft transplantation is a common spine procedure. Complications related to bone graft harvesting are still a major concern. Equine demineralized bone matrix has been reported to be osteoinductive. However, the application of equine bone protein extract in spine fusion has not been documented. In this experiment, we evaluated equine ...
Milner PI, Wilkins RJ, Gibson JS.To examine the effect of O(2) and the role, and source, of reactive oxygen species (ROS) on pH regulation in articular chondrocytes. Methods: Cartilage from equine metacarpo/tarsophalangeal joints was digested (collagenase) to isolate chondrocytes and loaded with 2',7'-bis-2-(carboxyethyl)-5(6)-carboxylfluorescein, a pH-sensitive fluorophore. O(2) tension was maintained using Eschweiler tonometers and a Wosthoff gas mixer. Cells were exposed to agents which alter ROS levels, mitochondrial inhibitors and/or inhibitors of protein phosphorylation. ROS levels were determined by dichlorofluorescein...
Rousseau K, Kirkham S, McKane S, Newton R, Clegg P, Thornton DJ.Horses frequently suffer from respiratory diseases, which, irrespective of etiology, are often associated with airway mucus accumulation. Studies on human airways have shown that the key structural components of the mucus layer are oligomeric mucins, which can undergo changes of expression and properties in disease. However, there is little information on these gel-forming glycoproteins in horse airways mucus. Therefore, the aims of this study were to isolate equine airways oligomeric mucins, characterize their macromolecular properties, and identify their gene products. To this end, pooled tr...
Aguilar PV, Weaver SC, Basler CF.Eastern equine encephalitis virus (EEEV) causes sporadic but often severe cases of human and equine neurological disease in North America. To determine how EEEV may evade innate immune responses, we screened individual EEEV proteins for the ability to rescue the growth of a Newcastle disease virus expressing green fluorescent protein (NDV-GFP) from the antiviral effects of interferon (IFN). Only expression of the EEEV capsid facilitated NDV-GFP replication. Inhibition of the antiviral effects of IFN by the capsid appears to occur through a general inhibition of cellular gene expression. For ex...
Luís C, Juras R, Oom MM, Cothran EG.There are three native Portuguese horse breeds: Lusitano, Sorraia and Garrano. This study compares diversity patterns of 17 protein and 12 microsatellite markers in these three as well as 30 other breeds to infer relationships among the breeds and to compare levels of polymorphism of these breeds for use in conservation efforts. The Garrano and the Lusitano showed a high level of genetic diversity, similar to that observed for most of the other analysed breeds, while the Sorraia and Friesian breeds showed low levels of variation for both genetic marker types. The combined protein and microsate...
Kölle S, Dubois CS, Caillaud M, Lahuec C, Sinowatz F, Goudet G.In the equine, the zona pellucida (ZP) is the major barrier to successful in vitro fertilization. Therefore the aim of our studies was to analyze species-specific features of the equine ZP in regard to structure and glycoprotein ZPB and ZPC expression sites during oocyte development and embryogenesis. The equine ZP revealed high immunological cross-reactivity to porcine ZPB and ZPC. In the ovary, the distribution of ZPB and ZPC was co-localized and correlated with the developmental stage of the follicle. ZPB and ZPC expression started in the oocyte of the late primordial and primary follicle. ...
Nakagawa K, Yamada Y, Fujiwara K, Ikeguchi M.Equine beta-lactoglobulin forms a compact intermediate at an acidic pH (A state). It also forms an expanded and helical conformation at low temperatures (C state). The structure of a single disulfide mutant C66A/C160A is similar to the A state in the presence of salts, while it is similar to the C state at low anion concentrations. We have investigated the temperature-dependent change in the secondary structure using circular dichroism and proline scanning mutagenesis. At low anion concentrations, the helical content increased linearly as temperature decreased. In the presence of salts, the A ...
Sayasith K, Doré M, Sirois J.The preovulatory rise in gonadotropins causes an expansion of the cumulus-oocyte complex, a process requiring the induction of several genes. The objectives of this study were to clone the equine tumor necrosis factor alpha-induced protein 6 (TNFAIP6), and investigate its regulation in equine follicles during human chorionic gonadotropin (hCG)-induced ovulation. The isolation of the equine TNFAIP6 cDNA revealed that it contains an open reading frame of 834 bp (including the stop codon), encoding a predicted 277 amino acid protein that is highly similar (91-93% identity) to known mammalian homo...
Krause JB, McKenzie HC.Adequate nutritional support of sick foals in critical care is an important aspect of treatment. When enteral feeding is contraindicated, parenteral nutrition (PN) provides a source of energy and protein. However, no study has critically assessed the use of PN in a large group of foals. Objective: The administration of PN to clinically ill foals was examined retrospectively to determine the effects of PN formulation and variables on the incidence of PN-associated complications and outcome. Objective: There was no effect of PN formula on 1) the occurrence or type of complications; 2) of PN form...
Elce YA, Orsini JA, Blikslager AT.To assess expression of cyclooxygenase (COX)-1 and -2 in naturally occurring squamous cell carcinomas (SCCs) and the analogous normal tissues in horses. Methods: Tissue samples collected from 3 conjunctival, 2 vulvar, 4 preputial, and 5 penile SCCs during surgical excision in 14 horses and from corresponding body regions (conjunctiva [n = 5 horses], vulva [2], prepuce [3], and penis [3]) in 5 horses euthanized for reasons unrelated to neoplasia. Methods: Tissue samples were snap frozen in liquid nitrogen and stored at -80 degrees C until analysis. Protein was extracted from the frozen tissues,...
al-Massad FK, Kadir FH, Moore GR.The origin of the 440 nm fluorescence of horse spleen ferritin and of Pseudomonas aeruginosa and Azotobacter vinelandii bacterioferritin has been investigated using a Nitro Blue Tetrazolium/glycinate colorimetric test specific for quiones [Paz, Flückiger, Boak, Kagan & Gallop (1991) J. Biol. Chem. 266, 689-692]. The results of the analysis indicate that ferritin and bacterioferritins contain quinones. A possible functional role of these quinones in iron uptake and release is described, as is the possibility that the presence of quinones in these proteins results from oxidative damage.
Stewart DR, Nevins B, Hadas E, Vandlen R.Relaxin, a polypeptide hormone normally associated with pregnancy, has been purified from many species, and the sequence determined for a growing number. Equine relaxin has been previously purified by acetone extraction, gel filtration, and ion exchange chromatographies. In an attempt to develop a more rapid and efficient method for relaxin purification, the use of affinity chromatography coupled with HPLC was explored. Monoclonal antibodies were raised against highly purified equine relaxin; large quantities of antibody were obtained by ascites production and attached to a solid phase support...
Klein C, Scoggin KE, Troedsson MH.Establishment of pregnancy is critically dependent upon a precisely orchestrated embryo-maternal interaction leading to a receptive uterine environment. The up-regulation of the interferon-stimulated protein 15 kDa (ISG15) during pregnancy has been described in various species and has been hypothesized to be part of the molecular repertoire that makes the uterus receptive to conceptus development. In the current study, the expression of ISG15 and enzymes involved in ISG15ylation was examined at the mRNA and protein level in equine endometrium at Day 14 of the luteal phase and at Day 14 and 50 ...
Clegg GA, Stansfield RF, Bourne PE, Harrison PM.An electron density map of horse spleen apoferritin at 0.28-nm (2.8 A) resolution and its preliminary interpretation have been described previously. Rigorous examination of this and newer maps at the same nominal resolution but calculated from more extensive data sets, including model building in a Richards' comparator, now allows us to report on structural features in more detail. We list inter-helical angles within and between neighbouring subunits, and describe a new short region of inter-subunit anti-parallel pleated sheet. A short section of electron density not properly accounted for in ...
Garcia-Suarez O, Germanà G, Naves FJ, Ciriaco E, Represa J, Vega JA.The medial wall of the vomeronasal organ (VNO) is lined with a sensory epithelium that is closely related to the olfactory epithelium, which is developed from the olfactory placode. It undergoes continuous replacement during its life span. In other sensory epithelia, cell proliferation is under the control of some trophic factors. Whether these proteins are involved in the continuous turnover of the VNO epithelium is unknown. This study approaches this topic by analyzing the occurrence of signal-transducing receptor proteins for neurotrophins (Trk proteins) and epidermal growth factor (EGFr). ...
Widders PR, Stokes CR, David JS, Bourne FJ.IgG, IgA, IgM and albumin concentrations were measured in serum, follicular fluid and oviductal, uterine and intestinal secretions of the horse. Follicular protein concentrations were found to be dependent on serum concentration and molecular size. Of the immunoglobulins only IgG was detectable in oviductal secretions, but IgG:albumin ratios did not differ significantly from those in serum. IgG, IgA and IgM were measured in uterine secretions, with IgG predominant. Serum transudation into uterine secretions was minimal. In intestinal secretions, IgA levels were slightly higher than IgG, with a...
Benton HP, Cheng TC, MacDonald MH.To determine the response of equine articular cartilage cells to heat and calcium stresses. Methods: Analysis of newly synthesized, [35S]methionine-labeled proteins after treatment of isolated primary equine chondrocytes. Methods: Primary cultures of equine articular chondrocytes were incubated at temperatures ranging from 37 to 42 C for heat stress experiments or incubated in the presence or absence of the intracellular calcium pump inhibitor, thapsigargin, for calcium stress experiments. Patterns of new protein synthesis were determined by incubating with [35S]methionine followed by separati...
Leisson K, Alev K, Kaasik P, Jaakma Ü, Seene T.This study investigates the myosin heavy chain (MyHC) isoform composition in the gluteus medius muscle of the Akhal-Teke horses using SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis). Fifteen horses aged between 1.5 and 23.5 years were used in this study and divided into three age groups: 1.5 to 4 (n = 6), 9 to 13 (n = 5) and 18.5 to 23.5 years (n = 4). The average content of the MyHC I isoform was 11.72 ± 1.07% (variation between individuals: 7.09% to 20.14%). The relative content of the MyHC IIa and IIx isoforms was subsequently 38.20 ± 1.46% (30.73% to 48.78%) and 50.0...
Peaker M, Rossdale PD, Forsyth IA, Falk M.Small samples of mammary secretion were taken for analysis from Thoroughbred mares during the last 3 weeks of pregnancy up to the time of foaling. The concentrations of sodium and chloride decreased while those of lactose, potassium, citrate, phosphate, calcium, magnesium and protein increased. The time-course of these changes showed marked variation between animals. The concentration of whey proteins began to increase about 10 days before parturition. The appearance of the secretion and the size of the mammary glands increased in the last few days of pregnancy. It is suggested that the concen...
Tydén E, Tjälve H, Larsson P.Among the cytochrome P450 enzymes (CYP), families 1-3 constitute almost half of total CYPs in mammals and play a central role in metabolism of a wide range of pharmaceuticals. This study investigated gene and protein expression and cellular localisation of CYP1A, CYP2A, CYP2C, CYP2D and CYP2E in equine intestine and liver. Real-time polymerase chain reaction (RT-PCR) was used to analyse gene expression, western blot to examine protein expression and immunohistochemical analyses to investigate cellular localisation. Results: CYP1A and CYP2C were the CYPs with the highest gene expression in the ...
Wilsher S, Gower S, Allen WR.A polyclonal human mucin-1 (MUC1) antibody was used to stain immunohistochemically for the presence of MUC1 on the endometrium and fetal membranes in mares between 20 and 309 days of gestation. Western blot analysis demonstrated the presence of a protein equivalent in size to a human MUC1 isoform, MUC1/Y, in equine endometrium, allantochorion and amnion. At all stages of gestation examined immunoreactivity to the MUC1 antibody was detected on the apical surface of the lumenal epithelium of the endometrium and the epithelium lining the mouths and apical regions of the endometrial glands. Furthe...
Cavalcanti MC, Rizgalla M, Geyer J, Failing K, Litzke LF, Bergmann M.In eukaryotic cells, the major protein constituents of the chromatin are histones, which can be divided into five classes, identified as H1, H2A, H2B, H3 and H4. During normal spermatogenesis, a testis-specific H1t is expressed in primary spermatocytes and believed to facilitate histone to protamine exchanges during spermiogenesis. In equine testes we detected the H1 protein at 22kDa by western blot analysis while H1t was detected at 29kDa. H1 protein was found to be expressed in all germ cells up to elongating spermatids (Sc) at stage IV. In peripubertal animals, there was a prolonged express...
Marycz K, Wiatrak B, Irwin-Houston JM, Marcinkowska K, Mularczyk M, Bourebaba L.Peroxisome proliferator-activated receptor gamma (PPARγ) is a transcription factor that promotes adipogenesis, lipid uptake and storage, insulin sensitivity, and glucose metabolism. Hence, defects in PPARγ have been associated to the development of metabolic disorders. Sex hormone-binding globulin (SHBG) is a glycoprotein primarily produced in the liver that regulates the bioavailability of sex hormones. Alike PPARγ, low SHBG levels have been correlated with insulin resistance and associated endocrine abnormalities. Therefore, this study aimed to verify whether SHBG may restore depleted PPA...
Patterson SD, Bell K.Three cases of abnormal expression of the equine protease inhibitory alleles, Pi F, L, and S1, were observed following the examination of 30,000 plasma samples by one-dimensional acid (pH 4.6) polyacrylamide gel electrophoresis. Characterization of the abnormal proteins in terms of isoelectric point, molecular mass, inhibitory spectra, and sialic acid content was performed using one- and two-dimensional electrophoretic techniques. The Pi F and S1 abnormalities were postulated to be the result of amino acid substitutions causing alterations in the processing of the carbohydrate side chains. No ...
Conti A, Godovac-Zimmermann J, Liberatori J, Braunitzer G.beta-Lactoglobulin-like proteins were detected in horse colostrum and normal milk using immunological techniques. In contrast to the beta-lactoglobulins sequenced so far these proteins are monomeric and genetically not homogenous. In this paper we report the first primary structure of a monomeric beta-lactoglobulin from horse colostrum. By means of an automatic liquid-phase sequenator the sequence of peptides obtained by tryptic digestion and by cyanogen bromide cleavage was determined. A limited tryptic digestion and hydrolysis with chymotrypsin provided the necessary overlapping peptides. Th...
Hunter DJ, Macmaster R, Roszak AW, Riboldi-Tunnicliffe A, Griffiths IR, Freer AA.Equine P2 protein has been isolated from horse spinal cord and its structure determined to 2.1 A. Since equine myelin is a viable alternative to bovine tissue for large-scale preparations, characterization of the proteins from equine spinal cord myelin has been initiated. There is an unusually high amount of P2 protein in equine CNS myelin compared with other species. The structure was determined by molecular replacement and subsequently refined to an R value of 0.187 (Rfree=0.233). The structure contains a molecule of the detergent LDAO and HEPES buffer in the binding cavity and is otherwise ...
McDowell KJ, Sharp DC, Fazleabas AT, Roberts RM.Conceptuses were obtained from pony mares on each day of pregnancy between Days 12 and 28, and on Days 39, 45, 65 and 100. Endometrium was obtained from mares at Days 12, 14, 16, 18, 39, 45, 65 and 100 of pregnancy, and from non-pregnant mares during anoestrus, during transition into the breeding season, at oestrus, or during dioestrus. Tissues were incubated in vitro for 24 h with L-[3H]leucine. Proteins synthesized and released into the culture medium were analysed by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and fluorography. Conceptuses obtained before Day 14 after ovul...
Smith HT, Millett F.Spectrophotometric titrations of five singly modified horse heart ferricytochromes c, specifically (trifluoromethyl)phenylcarbamylated (CF3PhNHCO-) or trifluoroacetylated (CF3CO-) at lysines-13, -72, and -79, were carried out. The CF3PhNHCO-Lys-13, Lys-79, and CF3CO-Lys-79 derivatives all underwent alkaline isomerization with loss of the 695-nm band to low-spin species with an apparent pK of about 8.9, as did the unmodified cytochrome. However, modification of lysine-72 appeared to alter the reaction pathway since the CF3PhNHCO-Lys-72 derivative isomerized to a high-spin form with an apparent ...
Reilas T, Katila T.Uterine lavage fluids from postpartum and nonparturient mares were compared to determine when the normal secretory capacity of the postpartum uterus is restored. Lavage fluids were obtained from cyclic nonparturient mares on the second, fourth or fifth day of oestrus, and 3, 8, or 14 days after ovulation (seven mares/sampling day). Twelve intact postpartum mares were sampled 1 to 28 days postpartum (group A: 1, 6, 12 and 20; group B: 2, 8, 14 and 24; group C: 4, 10, 16 and 28 days postpartum; four mares/group). Three ovariectomized (OVX) postpartum mares were sampled as mares in group C. Sampl...
Adler DMT, Serteyn D, Franck T, Jørgensen E, Christophersen MT, Denwood M, Verwilghen DR.To compare the extent of inflammation and catabolic collagen response in the middle carpal joints (MCJs) of healthy horses following intra-articular injection of 2% lidocaine, 2% mepivacaine, lactated Ringer solution (LRS), or 0.1% methyl parahydroxybenzoate. 17 adult horses. In the first of 2 experiments, the left middle carpal joint (MCJ) of each of 12 horses was injected with 10 mL of 2% lidocaine (n = 3), 2% mepivacaine (3), or LRS (control; 6). After a 4-week washout period, the right MCJ of the horses that received lidocaine or mepivacaine was injected with 10 mL of LRS, and the right MC...
Klimas NG, Caldwell KE, Whitney PL, Fletcher MA.Membrane glycoproteins from horse, sheep, goat and bovine erythrocytes were solubilized and purified. These glycoproteins could be placed in three groups based on their degrees of glycosylation: The major bovine erythrocyte glycoprotein (BGII) had 77% sugar, the minor bovine glycoprotein (BGI) had 27% sugar and the others had approximately 50% sugar. Four of the glycoproteins aggregated in a uniform way in aqueous solution--one, BGII, did not. Four had similar subunit sizes of 25-34,000 daltons, but BGII was larger--55,000 daltons. Receptor functions (for plant and invertebrate lectins, antibo...
Eckert RE, Neuder LE, Bell JL, Trujillo JC, Jones SL.The goal of this study was to define the role for p38 mitogen-activated kinase (MAPK) in the signaling mechanism regulating pro-inflammatory cyclooxygenase (COX) gene expression in lipopolysaccharide (LPS)-activated equine leukocytes for the purposes of identifying novel targets for anti-inflammatory therapy in endotoxemic horses. The p38 MAPK has been shown to positively regulate inflammatory gene expression in human leukocytes and can be activated by a variety of stimuli including LPS, TNF-alpha, and IL-1. Activation-associated phosphorylated p38 MAPK has been implicated in the up-regulation...
Saastamoinen M, Särkijärvi S, Suomala H.Six Finnhorse mares were used in a digestibility trial, in which six typical horse diets were compared. The diets were: (A) haylage 100%; (B) hay 100%; (C) hay 70% and oats 30%; (D) hay 70% and soybean meal + oats 30%; (E) hay 70%, rapeseed meal + oats 30% and (F) hay 70 %, linseed meal + oats 30%. The trial was conducted according to an unbalanced 6 × 4 Latin square design with four 3-week experimental periods. The experimental period consisted of 17-day preliminary feeding which was followed by a 4-day total faecal and urine collection periods to evaluate N excretion. The digestibilities of...
Hatzel JN, Bouma GJ, Cleys ER, Bemis LT, Ehrhart EJ, McCue PM.Early pregnancy factor has been identified as a 10-kDa extracellular homolog of heat shock protein 10 (Hsp10). Hsp10 has been detected during early pregnancy in serum of mice, sheep, pigs, horses, cows, and humans by the rosette inhibition test. Hsp10 has also been associated with several neoplastic and autoimmune diseases. The goal of the present study was to determine if Hsp10 could be detected in the early equine embryo through the use of immunohistochemistry and quantitative real-time PCR. Additionally, analysis of systemically harvested peripheral blood mononuclear cells (PBMCs) from both...
Bhuyan AK, Udgaonkar JB.The unfolding kinetics of horse cytochrome c in the oxidized state has been studied at 10, 22, and 34 degreesC as a function of guanidine hydrochloride (GdnHCl) concentration. Rapid (millisecond) measurements of far-UV circular dichroism (CD) as well as fluorescence quenching due to tryptophan to heme excitation energy transfer have been used to monitor the unfolding process. At 10 degreesC, the decrease in far-UV CD signal that accompanies unfolding occurs in two phases. The unobservable burst phase is complete within 4 ms, while the slower phase occurs over tens to hundreds of milliseconds. ...
Pawlak EA, Geor RJ, Watts MR, Black SJ, Johnson PJ, Belknap JK.Hypoxia-inducible factor-1α (HIF-1A) is an important protein in the regulation/induction of many genes in the cellular and tissue response to hypoxia and a central mediator in inflammatory signalling. As both hypoxia and inflammatory events are purported to occur in the lamellar epidermis in sepsis-related laminitis in the equid, HIF-1A may play a central role in this disease process. Objective: To assess the regulation of HIF-1A and HIF-1A-related genes in the equine keratinocyte in vitro and in the lamellar tissue of horses with sepsis-related laminitis. Methods: In vivo and in vitro experi...
Satterlee JD, Moench S.Proton hyperfine resonance assignments for cytochromes c from several species are currently being successfully pursued by several laboratories. These efforts focus mostly on the ferrous forms. In contrast to that work, we have pursued assignments of the proton hyperfine shifted resonances for horse and tuna ferricytochromes c. Our results indicate that assignments are nearly identical in those two proteins. Using the pre-steady state nuclear Overhauser effect, several additional assignments have been made for the tuna protein, whereas for the horse protein, the following protons have been assi...
Main AR, Soucie WG, Buxton IL, Arinc E.A relatively simple method is described by which cholinesterase was purified about 19000-fold starting from horse serum. Typically 20 litres of serum were processed to yield 15-18mg of electrophoretically pure cholinesterase in the form of an active salt-free dry powder. The method included two stages: fractionation with (NH(4))(2)SO(4) and ion-exchange chromatography. The (NH(4))(2)SO(4) stage included, in principle, the acid (pH3) step of the Strelitz (1944) procedure. The step took advantage of the stabilizing effect that 33%-satd. (NH(4))(2)SO(4) has on cholinesterase activity at pH3 and i...
Choi Y, Yoon M.Androstenone is the first mammalian steroidal pheromone to be identified. Pheromones are chemicals that animals use to communicate within a species. Pheromone detections are related to vomeronasal organ (VNO) and olfactory epithelium (OE) in mammals. Olfactory Receptor Family 7 Subfamily D Member 4 (OR7D4) is an odorant receptor that responds to androstenone. Several studies indicated that spray with androstenone changes behaviors of the boar and dogs. However, the expression of OR7D4 in VNO and OE was not reported in mammals except human. Thus, the main objectives of this study were to invest...
Magdesian KG.Horses with GI diseases such as colic and diarrhea are often intolerant of adequate enteral nutrition. Nutritional intervention should be an early part of therapeutic management in such cases. Protein and energy malnutrition in critically ill horses can have deleterious effects, including poor wound or incisional healing, reduced immunity, and weight loss. Early enteral or parenteral support should be provided to supply resting DE requirements in the equine ICU.
