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Topic:Protein
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.
Effects of Dietary Non-Fibrous Carbohydrate to Neutral Detergent Fiber Ratio on Apparent Digestibility, Fecal Microbiota, and Plasma Metabolomics in Yili Horses. This study aimed to investigate the effects of dietary NFC/NDF ratio on nutrient apparent digestibility, fecal fermentation parameters, microbial diversity, and plasma metabolomics in Yili horses. Twenty-four healthy Yili horses with similar body weights (406 ± 22.73 kg) were divided into four groups, each with six replicates: the Control Group (CG), Low-NFC Group (LG), Medium-NFC Group (MG), and High-NFC Group (HG). The experiment lasted 52 d, comprising a 7-day adaptation period and a 45-day experimental period. Total fecal collection was conducted from days 41 to 45 to calculate nutrient a...
Horse immune response of recombinant Lawsonia intracellularis vaccine: Assessing the immunomodulatory impact of probiotic supplementation. Equine proliferative enteropathy (EPE), caused by Lawsonia intracellularis induces intestinal epithelial hyperplasia, resulting in malabsorption, weight loss, colic, and diarrhea. Despite its clinical and economic importance, no equine-specific vaccine is commercially available. Recombinant subunit vaccines are a safe and scalable alternative; however, their immunogenicity often requires improvement. Probiotic supplementation with Bacillus toyonensis may act as an immunomodulatory strategy to potentiate vaccine efficacy. This study aimed to evaluate the immunogenicity of a recombinant protein ...
Interleukin-4 changes the transcriptome, ECM-associated components and function of mare endometrial fibroblast: Insights from healthy and fibrotic cells. Fibrosis remains incompletely understood, particularly in terms of how immune mediators shape stromal programs. We used a spontaneous large‑animal model-endometrosis (equine endometrial fibrosis) to define how interleukin‑4 (IL‑4) reprograms fibroblasts from healthy and fibrotic endometrium. Primary fibroblasts were exposed to IL‑4 (10 ng/mL) for 48 or 96 h. At 48 h, bulk transcriptomes revealed 1307 differentially expressed genes (DEGs; 648 up, 659 down) and 1271 DEGs (645 up, 626 down) in fibroblasts derived from endometria without or with endometrosis, respectively. Enrichment an...
Analysis of the antitrypsin activity and immunoglobulin G concentration in equine colostrum and milk. Immunoglobulins ingested via colostrum must remain intact to be absorbed by the neonate. Equine colostrum contains antitrypsin activity, which likely protects these proteins from proteolysis in the gastrointestinal tract. Objective: To quantify antitrypsin activity in equine colostrum and milk, describe its temporal changes during the first five days after parturition, compare actitvity between left and right mammary glands, evaluate differences between nulliparous and pluriparous mares, and examine associations with immunoglobulin G (IgG) concentrations and mare age. Methods: This retrospecti...
Genome-wide CRISPR/Cas9 screening identifies host factors critical for antiviral defense against equine herpesvirus type 1. Equine herpesvirus type 1 (EHV-1) is a major veterinary pathogen causing significant economic losses in the livestock industry. Despite its impact, effective vaccines and targeted antiviral strategies remain limited, largely due to an incomplete understanding of host factors regulating viral replication and pathogenesis. Unassigned: To systematically identify host genes essential for EHV-1 infection, we established a BHK-21 cell line stably expressing Cas9 and performed a genome-wide CRISPR/Cas9 knockout screen using a pooled lentiviral single-guide RNA library. Significantly enriched candidat...
Differential Expression and Analysis of TBX3 Gene in Skin Tissues of Dun Mongolian Horses with and Without Bider Markings. (1) Background: The dun coat color, a wild-type phenotype in horses characterized by pigment dilution and primitive markings, is regulated by . This study explored the expression and localization of in the Bider marking (a primitive mark unique to the shoulder of horses); (2) Methods: We compared skin tissues from Bider-marked and non-Bider dun Mongolian horses. Samples were collected from the Bider area (dark-colored/light-colored shoulder), dorsal midline, and croup. Histological staining, qRT-PCR, and Western blotting were used to analyze pigment distribution and expression at mRNA and pr...
Cell Surface Vimentin Is an Attachment Factor That Facilitates Equine Arteritis Virus Infection In Vitro. Our laboratory identified the susceptible allelic variant of equine CXCL16 protein (EqCXCL16S) as an entry receptor for equine arteritis virus (EAV). However, EAV has a broad host cell tropism and infects cells that lack EqCXCL16S. Thus, we hypothesized that EAV interacts with other host cell protein(s) that facilitate EAV infection. A virus overlay protein-binding assay in combination with a Far-Western blot from EAV-susceptible equine pulmonary artery endothelial cells (EECs) and equine dermal fibroblasts (E. Derm) identified a 57 kDa protein, present in the membrane fraction of the protein ...
Production and characterization of rNGFSP: a recombinant fusion immunogen eliciting dual anti-NGF and anti-Substance P therapeutic antibodies for Degenerative Joint Disease. Anti-NGF monoclonal antibodies have recently been approved for treating degenerative joint disease, including osteoarthritis pain, in dogs and cats. However, their widespread use is limited by high cost and the requirement for repeated injections. Nerve Growth Factor and Substance P play central roles in the initiation and maintenance of inflammation and chronic pain in OA. There is a pressing need for new, safe, cost-effective therapies that target the underlying mechanisms of OA chronic pain. Here, we designed and produced a novel recombinant fusion protein, termed rNGFSP, which functions as...
The Effect of Training on the Expression of Protein and Metabolites in the Plasma Exosomes of the Yili Horse. The effects of physical exercise on protein expression and metabolites in the plasma exosomes of horses are not yet clear. This study aims to elucidate the effects of exercise training on the expression of plasma extracellular vesicle proteins and metabolites in horses through multi-omics analysis, providing reference indicators for the training and performance evaluation of horses. Methods: Eight healthy 3-year-old horses were selected for the study, with four untrained horses as the control group and four trained horses as the training group. After conducting proteomic and metabolomic an...
High glucose in the extender impacts the metabolic phosphoproteome and modifies the phosphorylation of AKAP4 in stallion spermatozoa. Commercial extenders for equine semen typically contain high glucose concentrations; however, recent research indicates that these supraphysiological concentrations of glucose may be detrimental. Spermatozoa lack translational activity and depend on post-translational protein modifications for their regulation. Supraphysiological glucose concentrations are known to modify cell regulation, and we hypothesize that post-translational modification deregulation associated with supraphysiological glucose concentrations occurs in spermatozoa extended in high glucose media; and increasing the concentr...
Genome-Wide Characterization of the HOX Gene Family: Evolution and Expression Patterns in Donkey. The gene family plays an indispensable role in regulating embryonic development, cell differentiation, and morphogenesis. This study employed bioinformatics approaches for systematic analysis, ultimately identifying 33 gene family members from the donkey genome. Physicochemical property analysis revealed that the number of amino acids encoded ranged from 94 to 444, with 31 members classified as alkaline proteins. Their secondary structure was predominantly composed of random coils and alpha helices, and all members were localized to the nucleus. Conserved motif analysis further demonstrated ...
Sodium-glucose transporters SGLT1 and SGLT2 in equine renal, hepatic and pancreatic tissue. Insulin dysregulation is a hallmark of equine metabolic syndrome (EMS), and in recent years, pharmacological treatment with sodium-dependent glucose transporter 2 inhibitors (SGLT2i) have shown promise in reducing the risk of hyperinsulinemia-associated laminitis in horses diagnosed with EMS. In humans and laboratory animals, this transporter protein is responsible for the majority of renal tubular glucose reabsorption, however, the presence of this and the related sodium-dependent glucose transporter 1 (SGLT1) have not yet been studied in equine kidneys. Additionally, studies in humans and la...
Age-induced gene expression in Thoroughbred horse skeletal muscle highlights genes that enhance muscle architecture and function. Early skeletal muscle development is critical for young racehorses, yet research on the transcriptional changes during this period is limited. Additionally, the impact of age on the transcriptional response to exercise training in equine athletes is not well understood. A transcriptome-wide analysis of differential gene expression in skeletal muscle was performed for five untrained Thoroughbred horses sampled at rest at two years old (UR2) and three years old (UR3). A total of 136 differentially expressed genes (DEGs) were identified, with 95 increased and 41 decreased in expression. GO enrich...
Equine monocyte-derived macrophages revisited: isolation and comprehensive characterization of pro- versus anti-inflammatory polarisation. Macrophages play key roles in tissue homeostasis and regeneration-associated inflammation. Unlike humans, a reliable protocol to obtain and polarise equine monocyte-derived macrophages is lacking. In this study the polarisation of equine macrophages, derived from CD172a peripheral blood monocytes is described. After differentiation, IFN-γ/LPS or IL-4 were used to induce pro- and anti-inflammatory phenotypes, respectively. Evaluation criteria included morphology, mRNA (RT-qPCR) and protein expression (flow cytometry, immunofluorescence), nitric oxide and arginase production, cytokine secretion...
Equine oviduct-specific glycoprotein is modulated by hormones and sperm cells. Oviduct glycoprotein 1 (OVGP1) is a key protein involved in oviductal functions. β-estradiol (E) and progesterone (P), oxytocin (OXT) and tumor necrosis factor-α (TNFα) modulate the equine oviduct function, through prostaglandin regulation. The objective was to evaluate OVGP1 expression within each equine oviduct segment (infundibulum, ampulla isthmus), throughout the estrous cycle. The in vitro effect of (i) E, P, OXT, TNFα; and (ii) spermatozoa, on oviduct OVGP1 transcription and secretion was studied. Gene transcription was assessed by real-time PCR; protein expression by western blot; ...
Sequential changes in serum protein profiles in pregnant Spanish purebred mares. This study investigates sequential changes in serum protein fractions during gestation in 31 pregnant Spanish Purebred mares compared to 15 non-pregnant control. Total serum proteins (TSP) were measured using the Biuret method, while albumin (ALB), total globulins (tGLOB), and their subfractions (α₁, α₂, β₁, β₂, γ-GLOB) were analyzed by agarose gel electrophoresis. Pregnant mares showed significant changes in serum protein profiles throughout gestation. TSP and tGLOB concentrations increased from mid to late pregnancy (months 6-11), while ALB concentrations declined between months...
Pilot study of β-endorphin concentrations in horses with pituitary pars intermedia dysfunction using a newly validated enzyme-linked immunosorbent assay. β-endorphin, a proopiomelanocortin (POMC)-derived peptide secreted by pars intermedia melanotropes, may play a significant but underexplored role in pituitary pars intermedia dysfunction (PPID) pathophysiology and diagnosis. This study aimed to (1) validate a commercially available human β-endorphin enzyme-linked immunosorbent assay (ELISA) kit for equine samples, and (2) compare β-endorphin concentrations between horses with PPID and healthy controls. Assay validation included the generation of standard curves using purified synthetic equine β-endorphin and human β-endorphin standards, w...
Towards understanding mare endometrosis: ex vivo study on the role of relaxin. Mare endometrosis remains a poorly understood pathological process. Objective: Mare endometrial tissue with endometrosis was used to determine the effects of relaxin (RLX). Healthy tissues that were left untreated (H), or treated with vehicle (Hveh), and tissues with endometrosis that were left untreated (E), treated with vehicle (Eveh), and treated with RLX (10, 25, 50 nM; ER) were used for an ex vivo system for 72 h. Methods: Tissue histological examination, and immunoenzymatic measurement of the concentrations of transforming growth factor β (TGF-β), interleukins (IL-6 and IL-8), and pr...
Free Plasma Amino Acid Concentrations in Horses Fed Different Dosing Regimens of Hydrolysed Collagen. Hydrolysed collagen is used as a supplement for horses with osteoarthritis, hoof horn growth problems, and gastric ulcers. To determine the oral availability of a specific hydrolysed collagen supplement and the appropriate dose, six Warmblood mares were fed two different concentrations of the supplement: 100 g HC (C), 50 g HC (C), and a control of 0 g HC (C) during one week in a randomised cross-over design. On day 7, 14 and 21, blood sampling for amino acid (AA) analysis was performed, just prior to feeding the supplement (t = 0) and every hour after feeding for 8 h (t = 1-8). Statistical ana...
Neutralisation of the Immunoglobulin-Cleaving Activity of Streptococcus equi Subspecies equi IdeE by Blood Sera from Ponies Vaccinated with a Multicomponent Protein Vaccine. subspecies () is the cause of strangles, one of the most prevalent diseases of horses worldwide. The disease is characterised by fever and the formation of abscesses in the lymph nodes of the head and neck, which can restrict the airway. A multicomponent subunit vaccine, Strangvac, has been shown to effectively reduce clinical signs of strangles and to reduce its incidence. The aim of this study was to determine the immune response against the immunoglobulin-cleaving endopeptidase IdeE, a key protective component within the vaccine and the ability of antibodies to neutralize the proteolytic...
Molecular Mechanisms Underlying Differences in Athletic Ability in Racehorses Based on Whole Transcriptome Sequencing. This study aimed to compare blood samples from Yili horses with outstanding and average performance in 5000 m races through transcriptome sequencing, identify key differentially expressed genes, lncRNAs, and circRNAs, as well as related enriched pathways, and elucidate their regulatory networks. This study used six healthy four-year-old Yili stallions as subjects, divided into an excellent group (E group, = 3) and an ordinary group (O group, = 3) based on their 5000-m race performance. Blood RNA-Seq technology was used to analyze differentially expressed mRNAs, lncRNAs, and circRNAs. A total...
Proteomic profiling of endometrial tissue and plasma from mares with endometrial fibrosis†. Endometrial fibrosis leads to subfertility when periglandular fibrosis impairs endometrial gland function. Identifying distinct protein expressions in mares suffering from this condition is essential for understanding the pathogenesis and developing treatments to delay fibrotic progression. To gain a better understanding of protein dynamics underlying this condition, Data-Independent Acquisition Mass Spectrometry was utilized to comprehensively characterize and compare the equine endometrial and plasma proteome from mares with different severities of endometrial fibrosis. Mares in diestrus wer...
The Influence of Horse Age, High-Pressure Technique and Various Heat Treatment Methods on the Quality of Horse Meat. The aim of this study was to demonstrate the effect of horse age, high-pressure cooking, and various heat-treatment methods on the quality of horse meat. The research material consisted of samples of the longissimus thoracis muscle obtained from 14 horse carcasses from two age groups. Samples of the muscle were subjected to traditional cooking (TC), sous-vide cooking (S-V), high-pressure cooking (HHP), HHP + TC, and HHP + S-V. The chemical composition, physicochemical properties, color parameters, pigment levels, texture parameters, and sensory properties of the meat were determined. Exposing...
Commercial cytokine assay on equine cerebrospinal fluid does not distinguish equine degenerative myeloencephalopathy from cervical vertebral stenotic myelopathy. To measure and compare CSF cytokine concentrations among horses with equine neuroaxonal dystrophy (eNAD)/equine degenerative myeloencephalopathy (EDM), horses with cervical vertebral stenotic myelopathy (CVSM), and control horses and to evaluate for associations with clinical parameters. Unassigned: Banked equine CSF samples from horses with neurologic disease that underwent a complete neurologic examination and postmortem diagnosis confirmation of CVSM or eNAD/EDM or from control horses were included. Cytokines were measured with an equine-specific cytokine/chemokine magnetic bead multiplex p...
Comparative analysis of blood whole transcriptome profiles in Yili horses pre- and post-5000-meter racing. This study employed Yili horses participating in a 5000-meter race as a model to investigate exercise-induced gene expression alterations in peripheral blood using whole transcriptome sequencing. Jugular vein blood samples from the three leading horses were collected pre- and immediately post-race, yielding 2,171 differentially expressed mRNAs (2,080 upregulated, 91 downregulated), 4,375 differentially expressed LncRNAs (4,354 upregulated), and 68 differentially expressed circRNAs (64 upregulated). GO/KEGG analyses demonstrated significant enrichment of differential mRNAs in transmembrane tran...
Horse model of spontaneous atrial fibrillation share proteomic changes with humans. Horses and humans are among the few mammals susceptible to spontaneous atrial fibrillation (AF), both suffering from high recurrence rates after treatment. Treatment resistance is often attributed to progressive atrial remodeling, but current treatment options fail to effectively address this aspect. Here, we introduce a novel horse model of spontaneous AF to investigate the biological pathway changes in early stages of the disease. Through data-independent acquisition mass spectrometry on biopsies from the right and left atrium and left ventricular chamber of horses with early-stage persisten...
Exploratory Study of the Urine Protein-to-Creatinine Ratio in Apparently Healthy Horses. Currently, there is relatively little published information on the urine protein-to-creatinine (UPC) ratio in horses, a parameter commonly used in dogs and cats. The aim of this study was to determine the reference range of UPC in urine samples of apparently healthy horses and to assess the influence of age and sex. Urinalysis and the determination of urinary protein and creatinine concentrations and ratios were performed. A total of 118 urine samples from apparently healthy horses were included in the study: 10 from foals younger than 6 months and 108 from horses older than 1 year. The median...
Cytoplasmic and nuclear Survivin expression in healthy gastric mucosae of equids: A comparative study between horses, donkeys, and mules. Information regarding Survivin protein expression in the gastric mucosa of equids is scarce. This protein has been associated with functions related to modulating apoptosis and promoting mitosis in epithelial cells and is considered part of gastric cytoprotection mechanisms in humans and mice, maintaining mucosal integrity and regulating cellular renewal. Objective: This study aimed to determine the expression of cytoplasmic and nuclear Survivin in gastric mucosae of equids (horses, donkeys, and mules). Methods: A total of 30 healthy stomachs, 10 from each species, were evaluated using histolo...
Probing Wnt pathway and functional signal in equine melanocytic neoplasms through quantitative proteomics and immunohistochemistry. Equine melanocytic neoplasm (EMN) is a skin tumor commonly observed in grey horses. Limited research has yet to investigate proteomic profiles of EMN, particularly in the early stages and their expression patterns. This study, therefore, aimed to identify signature proteins from tissue biopsies to distinguish early EMN, severe EMN, and normal groups. Results: Using proteomic analysis of 19 tissue samples (normal: n = 6, early EMN: n = 7, severe EMN: n = 6) through LC-MS/MS, 12,310 proteins were identified. Differentially expressed proteins (DEPs) and functional interaction analysi...
Brain-derived neurotrophic factor and neurotrophic tyrosine receptor kinase-2 in stallion testes: insights into seasonal changes and potential roles in spermatogenesis. Brain-derived neurotrophic factor (BDNF) and its receptor neurotrophic tyrosine receptor kinase-2 (NTRK2) have known important roles in the central nervous system for neurite growth, survival, and differentiation. Nevertheless, the significance of BDNF in spermatogenesis remains unclear in stallions. Therefore, the present study was designed 1) to investigate the expression of BDNF and its receptor NTRK2 and 2) the seasonal variation in the expression patterns of BDNF and NTRK2 in stallions' testes. We used testes from eight postpubertal Thoroughbred stallions collected after a field castratio...
