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Topic:In Vitro Research

In vitro research involving horses refers to the study of equine cells, tissues, or biological molecules outside their normal biological context, typically in controlled laboratory environments. This research approach allows scientists to investigate cellular processes, molecular interactions, and the effects of various treatments without the ethical and logistical complexities of in vivo studies. In vitro studies contribute to understanding equine physiology, pathology, and pharmacology by providing insights into cellular responses to pathogens, drugs, and other stimuli. This page compiles peer-reviewed research studies and scholarly articles that explore various in vitro methodologies and their applications in equine science, including cell culture techniques, molecular assays, and drug efficacy testing.
Use of specific mitochondrial complex inhibitors to investigate mitochondrial involvement on horse sperm motility and ROS production.
Research in veterinary science    March 24, 2022   Volume 147 12-19 doi: 10.1016/j.rvsc.2022.03.017
Giaretta E, Mislei B, Martínez-Pastor F, Nesci S, Spinaci M, Galeati G, Nerozzi C, Mari G, Tamanini C, Bucci D.Equine spermatozoa highly rely on oxidative phosphorylation for their energy management. The present work aimed to characterize the role of mitochondria on horse sperm motility and ROS production by incubating spermatozoa with specific inhibitors of the different mitochondrial complexes. Equine spermatozoa were incubated 1 h and 3 h at 37 °C with: complex I inhibitor rotenone (5 μM, ROT), complex II inhibitor dimethyl-malonate (10 mM, DMM), complex III inhibitor antimycin A (1.8 μM, ANTI), the uncoupling agent carbonyl cyanide m-chlorophenyl hydrazine (5 μM, CCCP), ATP synthase inhi...
Theileria equi RAP-1a and RAP-1b proteins contain immunoreactive epitopes and are suitable candidates for vaccine and diagnostics development.
International journal for parasitology    March 19, 2022   Volume 52, Issue 6 385-397 doi: 10.1016/j.ijpara.2022.01.004
Onzere CK, Fry LM, Bishop RP, Da Silva M, Madsen-Bouterse SA, Bastos RG, Knowles DP, Suarez CE.Theileria equi is an obligate intracellular protozoan parasite that causes severe hemolytic anaemia in most equid species. Similar to other apicomplexan parasites, T. equi contains rhoptries whose contents have been implicated in host cell invasion and formation of the parasitophorous vacuole that is crucial for survival of the species within cells. Despite their importance, the composition of T. equi rhoptries and their role(s) in host cell invasion remain unexplored. To gain insight into these issues, we evaluated the expression, immunogenicity, and functional roles of two T. equi rhoptry-as...
Nanoforming Hyaluronan-Based Thermoresponsive Hydrogels: Optimized and Tunable Functionality in Osteoarthritis Management.
Pharmaceutics    March 17, 2022   Volume 14, Issue 3 doi: 10.3390/pharmaceutics14030659
Porcello A, Gonzalez-Fernandez P, Jordan O, Allémann E.Hyaluronic acid (HA) constitutes a versatile chemical framework for the development of osteoarthritis pain treatment by means of injection in the joints, so-called viscosupplementation. Without appropriate physico-chemical tuning, such preparations are inherently hindered by prompt in vivo degradation, mediated by hyaluronidases and oxidative stress. To prolong hydrogel residence time and confer optimized product functionality, novel thermoresponsive nanoforming HA derivatives were proposed and characterized. Combined use of sulfo-dibenzocyclooctyne-PEG4-amine linkers and poly(N-isopropylacryl...
Autologous conditioned serum in equine and human orthopedic therapy: A systematic review.
Research in veterinary science    March 15, 2022   Volume 146 34-52 doi: 10.1016/j.rvsc.2022.03.009
Tokawa PKA, Brossi PM, Baccarin RYA.This systematic review aims to compile and present information of studies evaluating the effectiveness of autologous conditioned serum (ACS) in the healing of tendon, ligament and articular lesions in humans and horses. A systematic search of articles using Medline, PubMed, Embase, Bireme and Google Scholar was conducted up to August 2020. Studies regarding ACS' use in human orthopedic lesions were included if classified as RCTs, cohort and case-controls. All studies regarding this therapy in equine medicine were included given their scarcity. Pre-clinical experimental studies were selected if...
Peptide Mediated Adhesion to Beta-Lactam Ring of Equine Mesenchymal Stem Cells: A Pilot Study.
Animals : an open access journal from MDPI    March 15, 2022   Volume 12, Issue 6 734 doi: 10.3390/ani12060734
Regenerative medicine applied to skin lesions is a field in constant improvement. The use of biomaterials with integrin agonists could promote cell adhesion increasing tissue repair processes. The aim of this pilot study was to analyze the effect of an α4β1 integrin agonist on cell adhesion of equine adipose tissue (AT) and Wharton's jelly (WJ) derived MSCs and to investigate their adhesion ability to GM18 incorporated poly L-lactic acid (PLLA) scaffolds. Adhesion assays were performed after culturing AT- and WJ-MSCs with GM18 coating or soluble GM18. Cell adhesion on GM18 containing PLLA sc...
Zona pellucida glycoproteins: Relevance in fertility and development of contraceptive vaccines.
American journal of reproductive immunology (New York, N.Y. : 1989)    March 15, 2022   Volume 89, Issue 2 e13535 doi: 10.1111/aji.13535
Gupta SK.Mammalian zona pellucida (ZP) is composed of three to four glycoproteins, which plays an important role during fertilization. Mutations in the genes encoding zona proteins are reported in women with empty follicle syndrome, degenerated oocytes and those with an abnormal or no ZP further emphasizing their relevance during fertility. Immunization with either native or recombinant ZP glycoproteins/proteins leads to curtailment of fertility in various animal species. Observed infertility is frequently associated with ovarian pathology characterized by follicular atresia and degenerative changes in...
Oxidative cleavage of cellulose in the horse gut.
Microbial cell factories    March 12, 2022   Volume 21, Issue 1 38 doi: 10.1186/s12934-022-01767-8
Liu N, Yu W, Guo X, Chen J, Xia D, Yu J, Li D.Lytic polysaccharide monooxygenases (LPMOs) belonging to the auxiliary activity 9 family (AA9) are widely found in aerobic fungi. These enzymes are O-dependent copper oxidoreductases that catalyze the oxidative cleavage of cellulose. However, studies that have investigated AA9 LPMOs of aerobic fungi in the herbivore gut are scare. To date, whether oxidative cleavage of cellulose occurs in the herbivore gut is unknown. Results: We report for the first time experimental evidence that AA9 LPMOs from aerobic thermophilic fungi catalyze the oxidative cleavage of cellulose present in the horse gut t...
Equine enterocytes actively oxidize l-glutamine, but do not synthesize l-citrulline or l-arginine from l-glutamine or l-proline in vitro.
Journal of animal science    March 12, 2022   Volume 100, Issue 4 skac077 doi: 10.1093/jas/skac077
Martinez RE, Leatherwood JL, Bradbery AN, Silvers BL, Fridley J, Arnold CE, Posey EA, He W, Bazer FW, Wu G.In livestock species, the enterocytes of the small intestine are responsible for the synthesis of citrulline and arginine from glutamine and proline. At present, little is known about de novo synthesis of citrulline and arginine in horses. To test the hypothesis that horses of different age groups can utilize glutamine and proline for the de novo synthesis of citrulline and arginine, jejunal enterocytes from 19 horses of three different age groups: neonates (n = 4; 7.54 ± 2.36 d of age), adults (n = 9; 6.4 ± 0.35 yr), and aged (n = 6; 22.9 ± 1.0 yr) with healthy gastrointestinal tracts were...
Technical Note: A method for determination of titanium dioxide concentration in fecal samples.
Journal of animal science    March 10, 2022   Volume 100, Issue 3 skac074 doi: 10.1093/jas/skac074
Fowler AL, Hayes SH, Crum AD, Lawrence LM.Titanium dioxide has been used as a marker for determining diet digestibility indirectly, but some authors have expressed difficulty in measuring TiO2 concentrations in fecal material. We developed an accurate and precise method to determine TiO2 concentrations in equine feces. The method includes dry-ashing samples, digestion with (NH4)2SO4 in concentrated sulfuric acid, followed by the addition of H2O2 to produce a yellow to orange color that can be read spectrophotometrically. Accuracy was tested by spike recovery, and precision was tested by examining the coefficient of variation (CV) betw...
Deep-horn Artificial Insemination With Frozen Thawed Semen After re-extension in Autologous Seminal Plasma May Improve Pregnancy Rates in Jennies.
Journal of equine veterinary science    March 10, 2022   Volume 112 103932 doi: 10.1016/j.jevs.2022.103932
Fanelli D, Tesi M, Monaco D, Diaz-Jimenez M, Camillo F, Rota A, Panzani D.In the literature, the very low pregnancy rates after artificial insemination (AI) with frozen semen in donkeys were improved in one study after re-extension of the frozen-thawed semen in autologous seminal plasma. The aims of our study were (1) to describe in vitro post-thaw parameters of donkey jackass semen after re-extension in seminal plasma (SP) or in INRA96 and (2) to compare pregnancy rates in jennies bred with frozen-thawed semen using two different AI protocols. Semen collected from two Amiata donkey stallions, known to be fertile, was frozen in INRA96 supplemented with 2% egg yolk, ...
Evaluation of the effect of conventional and natural antifungals on motility and kinetics of cooled stallion semen.
Reproduction in domestic animals = Zuchthygiene    March 9, 2022   Volume 57, Issue 7 701-710 doi: 10.1111/rda.14109
Usuga A, Gutiérrez V, López ME, Pérez LF, Jaramillo L, Rojano B, Restrepo G.Microbial growth in semen may cause a decline of sperm quality and fertility; however, the addition of antifungals to semen extender has been shown to impair the overall fertility of the sperm. The aim of this study was to evaluate the antifungal activity of conventional and natural compounds, and their effect on the motility and kinetics of cooled stallion semen. A total of 15 ejaculates from five stallions were collected using the artificial vagina. Each ejaculate was supplemented with: fluconazole at 12.5 (F1), 25 (F2) and 50 (F3) mg/ml; amphotericin-B at 6.5 (A1), 12.5 (A2) and 25 (A3) mg/...
146 Transcriptome characterisation of equine oocyte maturation.
Reproduction, fertility, and development    March 2, 2022   Volume 34, Issue 2 311 doi: 10.1071/RDv34n2Ab146
de la Fuente A, Scoggin C, Bradecamp E, Ali H, Troedsson M, Meyers S, Dini P.No abstract available
57 Influence of donor mare age on pre- and postimplanation embryo development within an equine ovum pick-up-intracytoplasmic sperm injection-embryo transfer (OPU-ICSI-ET) program over a three-year period.
Reproduction, fertility, and development    March 2, 2022   Volume 34, Issue 2 264 doi: 10.1071/RDv34n2Ab57
Lazzari G, Colleoni S, Barandalla M, Benedetti M, Duchi R, Galli C.No abstract available
73 Preimplantation genetic diagnosis of glycogen branching enzyme deficiency and sex determination in equine in vitro-produced embryos.
Reproduction, fertility, and development    March 2, 2022   Volume 34, Issue 2 272-273 doi: 10.1071/RDv34n2Ab73
Barandalla M, Colleoni S, Duchi R, Benedetti M, Perota A, Galli C, Lazzari G.No abstract available
94 Effect of glycine and creatine on the in vitro capacitation-related events in frozen/thawed equine sperm.
Reproduction, fertility, and development    March 2, 2022   Volume 34, Issue 2 284 doi: 10.1071/RDv34n2Ab94
Talbot SA, Diaz FA, Gutierrez-Castillo EJ, Walker CN, de Aguiar LH, Bondioli KR.No abstract available
35 The effect of different cooled storage protocols on frozen-thawed equine semen.
Reproduction, fertility, and development    March 2, 2022   Volume 34, Issue 2 252-253 doi: 10.1071/RDv34n2Ab35
van Heule M, Verstraete MH, Blockx Z, De Blende P, Dini P, Daels P.No abstract available
42 Comparison of three permeating cryoprotectant mixtures for equine immature oocyte vitrification.
Reproduction, fertility, and development    March 2, 2022   Volume 34, Issue 2 256 doi: 10.1071/RDv34n2Ab42
Angel-Velez D, De Coster T, Azari-Dolatabad N, Fernández-Montoro A, Benedetti C, Bogado Pascottini O, Van Soom A, Smits K.No abstract available
92 Extracellular vesicles from oviductal spheroids and uterine horn epithelial cells show different uptake times by equine spermatozoa and act upon capacitation.
Reproduction, fertility, and development    March 2, 2022   Volume 34, Issue 2 283 doi: 10.1071/RDv34n2Ab92
Lange-Consiglio A, Canesi S, Funghi F, Bosi G, Cremonesi F.No abstract available
The Semen Microbiome and Semen Parameters in Healthy Stallions.
Animals : an open access journal from MDPI    February 22, 2022   Volume 12, Issue 5 534 doi: 10.3390/ani12050534
Quiñones-Pérez C, Martínez A, Ortiz I, Crespo F, Vega-Pla JL.Despite the advances in reproductive technology, there is still a considerable number of low sperm quality cases in stallions. Recent studies in humans have detected several seminal microflora-spermatozoa associations behind some idiopathic infertility cases. However, no studies are available on horses, and there is limited information on the microflora present in stallion ejaculates. Accordingly, the objective of this study was to examine associations to the presence of bacteria families with five sperm quality parameters: concentration, total number of spermatozoa, total and progressive moti...
Effective Penetration of a Liposomal Formulation of Bleomycin through Ex-Vivo Skin Explants from Two Different Species.
Cancers    February 21, 2022   Volume 14, Issue 4 1083 doi: 10.3390/cancers14041083
Ferrari G, Pang LY, De Moliner F, Vendrell M, Reardon RJM, Higgins AJ, Chopra S, Argyle DJ.Bleomycin is a chemotherapy agent that, when administered systemically, can cause severe pulmonary toxicity. Bleosome is a novel formulation of bleomycin encapsulated in ultra-deformable (UD) liposomes that may be applicable as a topical chemotherapy for diseases such as non-melanoma skin cancer. To date, the ability of Bleosome to effectively penetrate through the skin has not been evaluated. In this study, we investigated the ability of Bleosome to penetrate through ex vivo skin explants from dogs and horses. We visualized the penetration of UD liposomes through the skin by transmission elec...
Tumor Cell Plasticity in Equine Papillomavirus-Positive Versus-Negative Squamous Cell Carcinoma of the Head and Neck.
Pathogens (Basel, Switzerland)    February 18, 2022   Volume 11, Issue 2 266 doi: 10.3390/pathogens11020266
Strohmayer C, Klang A, Kummer S, Walter I, Jindra C, Weissenbacher-Lang C, Redmer T, Kneissl S, Brandt S.Squamous cell carcinoma of the head and neck (HNSCC) is a common malignant tumor in humans and animals. In humans, papillomavirus (PV)-induced HNSCCs have a better prognosis than papillomavirus-unrelated HNSCCs. The ability of tumor cells to switch from epithelial to mesenchymal, endothelial, or therapy-resistant stem-cell-like phenotypes promotes disease progression and metastasis. In equine HNSCC, PV-association and tumor cell phenotype switching are poorly understood. We screened 49 equine HNSCCs for equine PV (EcPV) type 2, 3 and 5 infection. Subsequently, PV-positive versus -negative lesi...
Caudal foot placement superior to toe elevation for navicular palmaroproximal-palmarodistal-oblique image quality.
Equine veterinary journal    February 15, 2022   Volume 55, Issue 1 122-128 doi: 10.1111/evj.13563
Peeters MWJ, Thursby JJ, Watson HE, Berner D.Palmaroproximal-palmarodistal oblique (PaPr-PaDiO) radiographs are regularly obtained for a full evaluation of the navicular bone (NB). Despite their routine use, different acquisition techniques are described. Objective: To determine optimal foot placement and beam angle for obtaining PaPr-PaDiO views. Methods: In vitro experiment. Methods: A convenience sample of 26 disarticulated forelimbs were placed in six different positions using a leg press to mimic the weight-bearing position. In each position, navicular PaPr-PaDiO images were obtained with eight different beam angles. The resulting 1...
The Effects of Prostaglandin E2 Treatment on the Secretory Function of Mare Corpus Luteum Depends on the Site of Application: An in vivo Study.
Frontiers in veterinary science    February 15, 2022   Volume 8 753796 doi: 10.3389/fvets.2021.753796
Piotrowska-Tomala KK, Jonczyk AW, Szóstek-Mioduchowska AZ, Żebrowska E, Ferreira-Dias G, Skarzynski DJ.We examined the effect of prostaglandin (PG) E on the secretory function of equine corpus luteum (CL), according to the application site: intra-CL injection vs. an intrauterine (intra-U) administration. Moreover, the effect of intra-CL injection vs. intra-U administration of both luteotropic factors: PGE and human chorionic gonadotropin (hCG) as a positive control, on CL function was additionally compared. Mares were assigned to the groups ( = 6 per group): (1) an intra-CL saline injection (control); (2) an intra-CL injection of PGE (5 mg/ml); (3) an intra-CL injection of hCG (1,500 IU/ml); (4...
The type of anticoagulant used for plasma collection affects in vitro Rhodococcus equi assays.
BMC research notes    February 14, 2022   Volume 15, Issue 1 50 doi: 10.1186/s13104-022-05933-4
Rivolta AA, Pittman DC, Kappes AJ, Stancil RK, Kogan C, Sanz MG.The efficacy of Rhodococcus equi-specific hyperimmune plasma (HIP) is usually evaluated in vitro. Anticoagulants (AC) used for plasma collection can negatively impact bacterial replication but their effect on R. equi growth has not been evaluated. The aim of this study was to establish the effect that AC routinely used in veterinary medicine (ACD, KEDTA, Li Heparin, and Na Citrate) have on in vitro R. equi growth. To assess this, in vitro assays commonly used to test HIP efficacy (direct effect on microorganism and macrophage infection), were performed using each AC and non-treated bacteria. R...
Alginate encapsulation of stallion sperm for increasing storage stability.
Animal reproduction science    February 12, 2022   Volume 238 106945 doi: 10.1016/j.anireprosci.2022.106945
Pruß D, Oldenhof H, Wolkers WF, Sieme H.The aim of this study was to establish an alginate encapsulation procedure for stallion sperm, and investigate if sperm encapsulation enhances longevity during cold storage and survival after cryopreservation. First, biocompatibility of the compounds needed for encapsulation was tested and factors determining capsule structure were identified. Sperm encapsulation was realized either by depositing droplets (20 µL) of sperm solution supplemented with barium or calcium chloride (10 mM) in alginate solution (0.25%, w/v), or by adding sperm-alginate droplets in solution containing barium or calc...
The Induced Expression of BPV E4 Gene in Equine Adult Dermal Fibroblast Cells as a Potential Model of Skin Sarcoid-like Neoplasia.
International journal of molecular sciences    February 10, 2022   Volume 23, Issue 4 1970 doi: 10.3390/ijms23041970
Podstawski P, Samiec M, Skrzyszowska M, Szmatoła T, Semik-Gurgul E, Ropka-Molik K.The equine sarcoid is one of the most common neoplasias in the family. Despite the association of this tumor with the presence of bovine papillomavirus (BPV), the molecular mechanism of this lesion has not been fully understood. The transgenization of equine adult cutaneous fibroblast cells (ACFCs) was accomplished by nucleofection, followed by detection of molecular modifications using high-throughput NGS transcriptome sequencing. The results of the present study confirm that - and -mediated nucleofection strategy significantly affected the transcriptomic alterations, leading to sarcoid-like...
Ex Vivo and In Vitro Analysis Identify a Detrimental Impact of Neutrophil Extracellular Traps on Eye Structures in Equine Recurrent Uveitis.
Frontiers in immunology    February 10, 2022   Volume 13 830871 doi: 10.3389/fimmu.2022.830871
Fingerhut L, Yücel L, Strutzberg-Minder K, von Köckritz-Blickwede M, Ohnesorge B, de Buhr N.Equine recurrent uveitis (ERU) is a common ocular disease of horses and described as a model for human autoimmune uveitis. This immune-mediated, inflammatory condition progressively destroys the eye, ultimately leading to blindness. Genetic and autoimmune factors, next to infections with Leptospira, are discussed as key factors in the pathogenesis. Furthermore, a release of neutrophil extracellular traps (NETs) by activated neutrophils is involved. NETs are composed of decondensed chromatin and proteins that can immobilize invading pathogens. However, if NETs accumulate, they can contribute to...
Assessment of thrombin generation in horses using a calibrated automated thrombogram.
Veterinary clinical pathology    February 9, 2022   Volume 51, Issue 1 57-64 doi: 10.1111/vcp.13055
Leclere M, Chevalier Z, Dubuc V, Beauchamp G, Bédard C.The amount of thrombin generated reflects the endogenous thrombin potential (ETP), which depends on the balance of pro- and anticoagulant factors. The calibrated automated thrombogram (CAT) allows for the direct measurement of thrombin generation during the clotting process. Objective: (1) To describe the results of the CAT assay in horses, (2) to establish intra-assay and intra- and interindividual variation of thrombin generation in healthy horses, and (3) to compare in vitro low-molecular-weight heparin (LMWH) sensitivity between healthy and sick horses. The hypothesis for the last objectiv...
Fecal extract from obese horses induces an inflammatory response by murine macrophages in vitro.
American journal of veterinary research    February 2, 2022   Volume 83, Issue 5 419-425 doi: 10.2460/ajvr.21.02.0024
Roth P, Stanley J, Chamoun-Emanuelli A, Whitfield-Cargile C, Coleman M.To compare the inflammatory response of murine macrophages exposed to the enteric microbiome of obese horses versus nonobese horses. Methods: Fecal samples from 12 obese horses (body condition score ≥ 7/9) and 12 nonobese horses (body condition score 4 to 5/9) with similar dietary management. Methods: Fecal supernatant was prepared from frozen fecal samples. RAW 264.7 macrophage cells were exposed to the fecal extract. Inflammatory cytokine (interleukin-1β, tumor necrosis factor-α, and interleukin-6) gene expression was quantified via real-time quantitative reverse transcription PCR assay,...
First reported case of fragile foal syndrome type 1 in the Thoroughbred caused by PLOD1 c.2032G>A.
Equine veterinary journal    February 1, 2022   Volume 54, Issue 6 1086-1093 doi: 10.1111/evj.13547
Grillos AS, Roach JM, de Mestre AM, Foote AK, Kinglsey NB, Mienaltowski MJ, Bellone RR.Warmblood Fragile Foal Syndrome Type 1 (WFFS) is an autosomal recessive disorder reported previously only in warmbloods and thought to be caused by a variant in the gene procollagen-lysine,2-oxoglutarate 5-dioxygenase 1 (PLOD1, c.2032G>A, p.Gly678Arg). Given the presentation of this Thoroughbred case, we hypothesised that a similar genetic mechanism caused this phenotype. Objective: To describe the pathological and genetic findings on a foal presenting to a veterinary practice in the UK with skin lesions similar to other Ehlers-Danlos Syndromes, including those documented for warmbloods with W...
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