Guttural Pouch Mycosis: A Three-Step Therapeutic Approach.

The temporary ligation of the common carotid artery is performed as an emergency aid in cases of guttural pouch mycosis. Its usefulness is put into perspective after an anatomical summary of arterial vascularization involving a guttural pouch. It helps to better understand the need for the cranial (cerebral) and caudal (cardiac) occlusion of an arterial rupture by embolization in order to achieve maximum success in preventing and treating an hemorrhage. Topical oxygen therapy used alone or in a multimodal approach with embolization surgery is performed to promote healing of the inflammatory and mycotic lesions observed when an individual is affected. In conclusion, this three-step therapeutic approach should enable the equine practitioner to better orient their decision tree when faced with this condition which, while rare, can be potentially fatal if poorly treated.

Clinical findings, surgical techniques, prognostic factors for short-term survival and long-term outcome in horses with acquired inguinal hernias: Ninety-eight cases (2005-2020).

OBJECTIVE: To report findings, outcome and determine variables associated with survival in horses with acquired inguinal hernia (AIH). STUDY DESIGN: Retrospective study. ANIMALS: A total of 98 cases in 97 horses. METHODS: The medical records (2005-2020) of horses diagnosed with AIH were reviewed. Retrieved data included signalment, history, clinical variables, surgical aspects, postoperative complications, and short- and long-term outcomes. Logistic regression analyses were used to determine factors associated with short-term survival (p < .05). RESULTS: Manual reduction was attempted in a third of the cases (32/98, 33%) and emergency surgery to reduce the hernia was performed in 64 of 98 (65%) cases. Concurrent small intestinal (SI) volvulus was identified in 26 (26/98, 27%) cases. Castration was the most common technique used to prevent recurrence (64/94, 68%). Overall AIH recurrence rate was 11% (11/98). A total of 59 (59/98, 60%) cases survived to hospital discharge and 49 of 52 (94%) cases were still alive after 12 months. Cases admitted within 10 h of colic signs had increased odds of survival (72%) compared to those admitted after 10 h (26%; p < .001). Draft breeds (p = .021), high heart rate on admission (p = .001) and concurrent SI volvulus (p = .048) were associated with reduced survival to hospital discharge. CONCLUSIONS: Horses with AIH had a higher risk of concurrent SI volvulus and lower survival than reported. Draft breeds, high heart rate on admission and concurrent SI volvulus were associated with reduced short-term survival. CLINICAL SIGNIFICANCE: The results of this study should help in prognostication for horses with AIH.

Effect of Allogeneic Oral Mucosa Mesenchymal Stromal Cells on Equine Wound Repair.

OBJECTIVE: To assess the clinical value and safety of the application of allogeneic equine oral mucosa mesenchymal stromal cells (OM-MSCs) to wounds. Animals. 8 healthy adult horses without front limb skin lesions or musculoskeletal disease. Procedures. Stem cells were isolated from the oral mucosa of a donor horse. Horses were subjected to the creation of eight full-thickness cutaneous wounds, two on each distal forelimb (FL) and two on both sides of the thorax (TH). Each wound was subjected to one out of four treatments: no medication (T1), hyaluronic acid- (HA-) gel containing OM-MSC (T2), HA-gel containing OM-MSC secretome (T3), and HA-gel alone (T4). Gross macroscopic evaluation and laser digital photographic documentation were regularly performed to allow wound assessment including wound surface area. Full-thickness skin punch biopsy was performed at each site before wound induction (D0, normal skin) and after complete wound healing (D62, repaired skin). RESULTS: All wounds healed without adverse effect at D62. Distal limb wounds are slower to heal than body wounds. OM-MSC and its secretome have a positive impact on TH wound contraction. OM-MSC has a positive impact on the contraction and epithelialization of FL wounds. No significant difference between wound sites before and after treatment was noted at histological examination. Conclusion and Clinical Relevance. Using horse cells harvested from oral mucosa is a feasible technique to produce OM-MSC or its secretome. The gel produced by the combination of these biologic components with HA shows a positive impact when applied during the early stage of wound healing.

The Effect of Topical Oxygen Therapy in Horses Affected with Mycosis of the Guttural Pouch: An Experimental Pilot Study and a Case Series.

Background: The management of bleeding originating from the guttural pouch (GP) has a high success rate, but the resolution of the macroscopic inflammatory lesions in the case of mycosis (GPM) is highly variable; the resolution of neurological disorders is inconstant and challenging. Objectives: Our aim was to test the feasibility and safety of topical oxygen therapy (TOT) in horses after induction of GPM and in cases with naturally occurring disease. Study design: This study was an in vivo experimental and retrospective two-phase study. Methods: During phase 1, the pilot study, both GPs were inoculated with Aspergillus fumigatus. One GP was randomly assigned to receive one to four TOT 30 min sessions with 100% medical oxygen at 9 L/min. Follow-up endoscopic images were assessed for scoring macroscopic inflammatory lesions of the pharynx and both GPs. In phase 2, the clinical study, TOT was administered for 45 to 60 min at 15 L/min in six horses presenting with GPM. Results: In phase 1, TOT administration was easy to perform in the standing horse with no adverse effects. After more than two administrations, macroscopic inflammatory lesions decreased more quickly in size in the treated GP. In phase 2, horses were treated with TOT only (n = 1) or combined with a transarterial coil embolization (TACE) procedure (n = 5). After TOT and discharge from the hospital, nasal discharge resolved in three horses, and improvement was noted in the fourth one. Between days 2 and 10 after admission, upper respiratory tract endoscopy (URTE) indicated size reduction and alteration in the appearance of all the macroscopic inflammatory lesions. The partial or total recovery of neurological disorders (2/4 laryngeal hemiparesis, 3/5 dysphagia, 1/2 dorsal displacement of the soft palate (DDSP), and 1/1 Horner's syndrome) was recorded. Main limitations: In phase 1, the small number of horses did not allow for statistically significant conclusions; in phase 2, clinical signs at admission varied between horses, which made comparison difficult. Conclusions: In adult horses, TOT alone or in combination with TACE is feasible and safe with a propensity to reverse the course and the progression of inflammatory lesions without additional local or systemic treatment.

Ex Vivo Comparison of a UV-Polymerizable Methacrylate Adhesive versus an Inverting Pattern as the Second Layer of a Two-Layer Hand-Sewn Jejunal Anastomosis in Horses: A Pilot Study.

Resection and anastomosis of small intestine during colic can lead to adhesions and recurrent colic. Several methods are available to reduce the rate of adhesions in the postoperative period, such as the use of serosal barriers. Surgical glues form a smooth surface, are fast to apply, and could reduce surgery time when performing anastomosis. A recently developed UV-polymerizable methacrylate adhesive (UV-PMA) is designed to anchor into the biological tissues' top surface offering sealant and a smooth cover over the anastomosis site. This adhesive was used ex vivo on fifteen samples of equine jejunum as the second layer of a two-layer anastomosis (1L-UV-PMA group) and compared to a two-layer anastomosis (simple continuous pattern covered with a Cushing pattern; 2L-CT group), in terms of feasibility, bursting strength pressure (BSP), luminal diameter reduction (LDR), and time of construction. Data were analysed using a paired t-test or a chi2-test (P < 0.05). The results showed no statistical difference in BSP, LDR, or any mode of failure between the two anastomosis types. However, the glue anastomosis formed a tunnel-like anastomosis and shredded under pressure, before apparition of leakage, preventing its usage in clinical cases with this methodology. It was concluded that modification of the technique is warranted before testing in clinical cases. A preprint of a former version of the manuscript is available on researchsquare.com, which was not conducted to print and publication after peer reviewing. Since then, the manuscript has been modified to this current version.

Mesenchymal stem cell transplantation into the spinal cord of healthy adult horses undergoing cervical ventral interbody fusion.

OBJECTIVE: To determine the feasibility of umbilical cord-derived mesenchymal stem cell (UC-MSC) transplantation into the cervical spinal cord of horses by using fluoroscopy with or without endoscopic guidance and to evaluate the neurological signs and tissue reaction after injection. STUDY DESIGN: Experimental study. ANIMALS: Eight healthy adult horses with no clinical signs of neurological disease. METHODS: After cervical ventral interbody fusion (CVIF), ten million fluorescently labeled allogeneic UC-MSC were injected into the spinal cord under endoscopic and fluoroscopic guidance (n = 5) or fluoroscopic guidance only (n = 3). Postoperative neurological examinations were performed, and horses were humanely killed 48 hours (n = 4) or 14 days (n = 4) postoperatively. Spinal tissues were examined after gross dissection and with bright field and fluorescent microscopy. RESULTS: Needle endoscopy of the cervical canal by ventral approach was associated with intraoperative spinal cord puncture (2/5) and postoperative ataxia (3/5). No intraoperative complications occurred, and one (1/3) horse developed ataxia with cell transplantation under fluoroscopy alone. Umbilical cord-derived MSC were associated with small vessels and detected up to 14 days in the spinal cord. Demyelination was observed in six of eight cases. CONCLUSION: Fluoroscopically guided intramedullary UC-MSC transplantation during CVIF avoids spinal cord trauma and decreases risk of ataxia from endoscopy. Umbilical cord-derived MSC persist in the spinal cord for up to 14 days. Cell injection promotes angiogenesis and induces demyelination of the spinal tissue. CLINICAL SIGNIFICANCE: Umbilical cord-derived MSC transplantation into the spinal cord during CVIF without endoscopy is recommended for future evaluation of cell therapy in horses affected by cervical vertebral compressive myelopathy.

Experimental induction of mycotic plaques in the guttural pouches of horses.

Guttural pouch mycosis (GPM) is a rare but potentially life-threatening condition in horses. GPM is caused by a fungal invasion into the mucosal lining of the guttural pouches and, frequently, the associated neurovascular structures. Although several species of fungi have been associated with this disease, Aspergillus spp. appear to be the most common isolated from the guttural pouches. However, it remains unclear which are the predisposing factors leading to the development of the infection. The objectives of the present study were to experimentally reproduce an infection by Aspergillus fumigatus and to follow the natural evolution of the mycosis. Eight guttural pouches from four horses were experimentally infected by endoscopy-guided intrapouch inoculation of A. fumigatus culture. Horses were monitored for clinical signs and development of fungal plaques through endoscopic examination. Mycotic lesions were observed in all the horses and a spontaneous regression was observed within 15-28 days. No development of clinical signs was noticed. In conclusion, we were able to induce the development of mycotic lesions and to observe a natural regression of these lesions without clinical signs.

Utility of a Mouse Model of Osteoarthritis to Demonstrate Cartilage Protection by IFNγ-Primed Equine Mesenchymal Stem Cells.

OBJECTIVE: Mesenchymal stem cells isolated from adipose tissue (ASC) have been shown to influence the course of osteoarthritis (OA) in different animal models and are promising in veterinary medicine for horses involved in competitive sport. The aim of this study was to characterize equine ASCs (eASCs) and investigate the role of interferon-gamma (IFNγ)-priming on their therapeutic effect in a murine model of OA, which could be relevant to equine OA. METHODS: ASC were isolated from subcutaneous fat. Expression of specific markers was tested by cytometry and RT-qPCR. Differentiation potential was evaluated by histology and RT-qPCR. For functional assays, naïve or IFNγ-primed eASCs were cocultured with peripheral blood mononuclear cells or articular cartilage explants. Finally, the therapeutic effect of eASCs was tested in the model of collagenase-induced OA (CIOA) in mice. RESULTS: The immunosuppressive function of eASCs on equine T cell proliferation and their chondroprotective effect on equine cartilage explants were demonstrated in vitro. Both cartilage degradation and T cell activation were reduced by naïve and IFNγ-primed eASCs, but IFNγ-priming enhanced these functions. In CIOA, intra-articular injection of eASCs prevented articular cartilage from degradation and IFNγ-primed eASCs were more potent than naïve cells. This effect was related to the modulation of eASC secretome by IFNγ-priming. CONCLUSION: IFNγ-priming of eASCs potentiated their antiproliferative and chondroprotective functions. We demonstrated that the immunocompetent mouse model of CIOA was relevant to test the therapeutic efficacy of xenogeneic eASCs for OA and confirmed that IFNγ-primed eASCs may have a therapeutic value for musculoskeletal diseases in veterinary medicine.

LOW-FIELD MAGNETIC RESONANCE IMAGING APPEARANCE OF POSTARTHROSCOPIC MAGNETIC SUSCEPTIBILITY ARTIFACTS IN HORSES.

An awareness of magnetic susceptibility artifacts is important for interpreting prepurchase and postoperative magnetic resonance imaging (MRI) studies in horses. These artifacts occur when a metallic or a paramagnetic substance creates a local magnetic field deformity. Aims of the current experimental study were to determine prevalence of these artifacts after arthroscopy in a sample of nonlame horses, and to describe effects of time and type of pulse sequence on low-field MRI signal intensity and detection of the artifacts. Ten, nonlame Standardbred horses were prospectively recruited. All horses underwent arthroscopy of both metacarpophalangeal joints for purposes unrelated to the study. Serial low-Field MRI examinations were performed on each horse and each joint (before, and 6 and 12 weeks postsurgery). In two horses, more detailed longitudinal evaluations were performed with additional MRI examinations. Magnetic susceptibility artifacts were detected postoperatively at the surgical access sites in eight metacarpophalangeal joints at both 6 and 12 weeks after surgery (40% prevalence). Neither of the two longitudinally followed horses had artifacts at any time. Artifacts were only detected on gradient echo (GRE) sequences. Findings indicated that magnetic susceptibility artifacts can be present in postarthroscopy MRI studies in horses and can persist up to 12 weeks after arthroscopy. For this sample of horses, the artifacts did not interfere with evaluation of the joint. Further longitudinal studies are needed to determine the full duration of magnetic susceptibility artifact persistence in affected tissues.

Development of an equine groove model to induce metacarpophalangeal osteoarthritis: a pilot study on 6 horses.

The aim of this work was to develop an equine metacarpophalangeal joint model that induces osteoarthritis that is not primarily mediated by instability or inflammation. The study involved six Standardbred horses. Standardized cartilage surface damage or "grooves" were created arthroscopically on the distal dorsal aspect of the lateral and medial metacarpal condyles of a randomly chosen limb. The contralateral limb was sham operated. After 2 weeks of stall rest, horses were trotted 30 minutes every other day for 8 weeks, then evaluated for lameness and radiographed. Synovial fluid was analyzed for cytology and biomarkers. At 10 weeks post-surgery, horses were euthanized for macroscopic and histologic joint evaluation. Arthroscopic grooving allowed precise and identical damage to the cartilage of all animals. Under the controlled exercise regime, this osteoarthritis groove model displayed significant radiographic, macroscopic, and microscopic degenerative and reactive changes. Histology demonstrated consistent surgically induced grooves limited to non-calcified cartilage and accompanied by secondary adjacent cartilage lesions, chondrocyte necrosis, chondrocyte clusters, cartilage matrix softening, fissuring, mild subchondral bone inflammation, edema, and osteoblastic margination. Synovial fluid biochemistry and cytology demonstrated significantly elevated total protein without an increase in prostaglandin E2, neutrophils, or chondrocytes. This equine metacarpophalangeal groove model demonstrated that standardized non-calcified cartilage damage accompanied by exercise triggered altered osteochondral morphology and cartilage degeneration with minimal or inefficient repair and little inflammatory response. This model, if validated, would allow for assessment of disease processes and the effects of therapy.

Percutaneous ultrasound-guided arterial angiography for transarterial coil placement in anesthetized and standing horses.

OBJECTIVE: To (1) describe ultrasound-guided percutaneous introduction of a transarterial angiographic catheter into the common carotid artery (CCA); (2) investigate the feasibility of using angiography of the carotid arteries in the guttural pouch region and assess transarterial coil (TAC) placement into the internal carotid artery (ICA). STUDY DESIGN: Experimental study. ANIMALS: Healthy Standardbred horses (n = 6), aged 5-8 years. METHODS: Six horses had ultrasound-guided percutaneous CCA catheterization and angiography under general anesthesia. Catheterization sites were ultrasonographically evaluated postoperatively. Ten weeks later using the same horses sedated and standing, the same procedure was combined with placement of a TAC in the ICA. RESULTS: Agitated contrast ultrasonography confirmed successful catheterization of the CCA. Needle puncture and introducer-set penetration of the CCA were the main technical difficulties. Radiography and fluoroscopy confirmed successful angiography and TAC placement. Mild hematoma formation was recorded in 4 of 12 procedures. CONCLUSION: Angiography and TCA placement in the ICA can be safely performed using a percutaneous approach to the CCA under ultrasound guidance, in standing or anesthetized horses. This approach might be used for TAC embolization procedure; however, technical difficulties and hematoma formation can impair the procedure.

Omics technologies provide new insights into the molecular physiopathology of equine osteochondrosis.

BACKGROUND: Osteochondrosis (OC(D)) is a juvenile osteo-articular disorder affecting several mammalian species. In horses, OC(D) is considered as a multifactorial disease and has been described as a focal disruption of endochondral ossification leading to the development of osteoarticular lesions. Nevertheless, OC(D) physiopathology is poorly understood. Affected horses may present joint swelling, stiffness and lameness. Thus, OC(D) is a major concern for the equine industry. Our study was designed as an integrative approach using omics technologies for the identification of constitutive defects in epiphyseal cartilage and/or subchondral bone associated with the development of primary lesions to further understand OC(D) pathology. This study compared samples from non-affected joints (hence lesion-free) from OC(D)-affected foals (n = 5, considered predisposed samples) with samples from OC-free foals (n = 5) considered as control samples. Consequently, results are not confounded by changes associated with the evolution of the lesion, but focus on altered constitutive molecular mechanisms. Comparative proteomics and micro computed tomography analyses were performed on predisposed and OC-free bone and cartilage samples. Metabolomics was also performed on synovial fluid from OC-free, OC(D)-affected and predisposed joints. RESULTS: Two lesion subtypes were identified: OCD (lesion with fragment) and OC (osteochondral defects). Modulated proteins were identified using omics technologies (2-DE proteomics) in cartilage and bone from affected foals compare to OC-free foals. These were associated with cellular processes including cell cycle, energy production, cell signaling and adhesion as well as tissue-specific processes such as chondrocyte maturation, extracellular matrix and mineral metabolism. Of these, five had already been identified in synovial fluid of OC-affected foals: ACTG1 (actin, gamma 1), albumin, haptoglobin, FBG (fibrinogen beta chain) and C4BPA (complement component 4 binding protein, alpha). CONCLUSION: This study suggests that OCD lesions may result from a cartilage defect whereas OC lesions may be triggered by both bone and cartilage defects, suggesting that different molecular mechanisms responsible for the equine osteochondrosis lesion subtypes and predisposition could be due to a defect in both bone and cartilage. This study will contribute to refining the definition of OC(D) lesions and may improve diagnosis and development of therapies for horses and other species, including humans.

Cecal vascular hamartoma causing recurrent colic in an Arabian mare.

A 5-year-old mare was treated for recurrent colic and weight loss by surgical removal of an intraluminal cecal mass. Microscopic examination revealed vascular hamartoma. A 6-month follow-up showed an improvement in the general condition of the mare. Vascular hamartoma should be one of the differential diagnoses for weight loss and colic.

Coliques récurrentes causées par un hamartome vasculaire caecal chez une jument Arabe. Une jument de 5 ans a été traitée pour coliques récurrentes et perte de poids par exérèse chirurgicale d'une masse caecale intraluminale. L'examen microscopique a révélé un hamartome vasculaire. Un suivi de 6 mois a montré une amélioration de l'état général de la jument. L'hamartome vasculaire doit faire partie du diagnostic différentiel de l'amaigrissement et des coliques.(Traduit par les auteurs).

Molecular dynamics simulations and structure-based rational design lead to allosteric HCV NS5B polymerase thumb pocket 2 inhibitor with picomolar cellular replicon potency.

The design and preliminary SAR of a new series of 1H-quinazolin-4-one (QAZ) allosteric HCV NS5B thumb pocket 2 (TP-2) inhibitors was recently reported. To support optimization efforts, a molecular dynamics (MD) based modeling workflow was implemented, providing information on QAZ binding interactions with NS5B. This approach predicted a small but critical ligand-binding induced movement of a protein backbone region which increases the pocket size and improves access to the backbone carbonyl groups of Val 494 and Pro 495. This localized backbone shift was consistent with key SAR results and was subsequently confirmed by X-ray crystallography. The MD protocol guided the design of inhibitors, exploiting novel H-bond interactions with the two backbone carbonyl groups, leading to the first thumb pocket 2 NS5B inhibitor with picomolar antiviral potency in genotype (gt) 1a and 1b replicons (EC50 = 120 and 110 pM, respectively) and with EC50 ≤ 80 nM against gt 2-6.

Structure-based design of novel HCV NS5B thumb pocket 2 allosteric inhibitors with submicromolar gt1 replicon potency: discovery of a quinazolinone chemotype.

We describe the structure-based design of a novel lead chemotype that binds to thumb pocket 2 of HCV NS5B polymerase and inhibits cell-based gt1 subgenomic reporter replicons at sub-micromolar concentrations (EC50<200nM). This new class of potent thumb pocket 2 inhibitors features a 1H-quinazolin-4-one scaffold derived from hybridization of a previously reported, low affinity thiazolone chemotype with our recently described anthranilic acid series. Guided by X-ray structural information, a key NS5B-ligand interaction involving the carboxylate group of anthranilic acid based inhibitors was replaced by a neutral two-point hydrogen bonding interaction between the quinazolinone scaffold and the protein backbone. The in vitro ADME and in vivo rat PK profile of representative analogs are also presented and provide areas for future optimization of this new class of HCV polymerase inhibitors.

Discovery and structural characterization of a new inhibitor series of HIV-1 nucleocapsid function: NMR solution structure determination of a ternary complex involving a 2:1 inhibitor/NC stoichiometry.

The nucleocapsid (NC) protein is an essential factor with multiple functions within the human immunodeficiency virus type 1 (HIV-1) replication cycle. In this study, we describe the discovery of a novel series of inhibitors that targets HIV-1 NC protein by blocking its interaction with nucleic acids. This series was identified using a previously described capsid (CA) assembly assay, employing a recombinant HIV-1 CA-NC protein and immobilized TG-rich deoxyoligonucleotides. Using visible absorption spectroscopy, we were able to demonstrate that this new inhibitor series binds specifically and reversibly to the NC with a peculiar 2:1 stoichiometry. A fluorescence-polarization-based binding assay was also developed in order to monitor the inhibitory activities of this series of inhibitors. To better characterize the structural aspect of inhibitor binding onto NC, we performed NMR studies using unlabeled and (13)C,(15)N-double-labeled NC(1-55) protein constructs. This allowed the determination of the solution structure of a ternary complex characterized by two inhibitor molecules binding to the two zinc knuckles of the NC protein. To the best of our knowledge, this represents the first report of a high-resolution structure of a small-molecule inhibitor bound to NC, demonstrating sub-micromolar potency and moderate antiviral potency with one analogue of the series. This structure was compared with available NC/oligonucleotide complex structures and further underlined the high flexibility of the NC protein, allowing it to adopt many conformations in order to bind its different oligonucleotide/nucleomimetic targets. In addition, analysis of the interaction details between the inhibitor molecules and NC demonstrated how this novel inhibitor series is mimicking the guanosine nucleobases found in many reported complex structures.

Trans-arterial coil embolization of the internal carotid artery in standing horses.

OBJECTIVES: To develop transarterial coil embolization (TACE) for occlusion of the internal carotid artery (ICA), in normal standing horses, and to evaluate it use for prevention of hemorrhage in horses with guttural pouch mycosis (GPM). STUDY DESIGN: Prospective study. ANIMALS: Normal horses (n = 8) and 5 with GPM. METHODS: Horses had TACE of the ICA in standing position under fluoroscopic guidance. Four normal horses were euthanatized 2 weeks after TACE for morphologic assessment and 4 were followed for 6 months. The 5 clinically affected horses were evaluated for long-term (10-12 months) success rate and complications. RESULTS: No complications related to the TACE were noted. Up to 30 mL warmed meglumine ioxithalamate was injected and well tolerated. Standing angiography confirmed complete occlusion of all vessels, and coils were positioned as intended; the procedure did not alter local hemodynamics. At 2 weeks, maturing to mature continuous thrombi was seen at the site of the coils. Two clinically affected horses died at day 3 and 12 after surgery from other problems. In the 3 surviving horses, mycotic lesions completely resolved without additional treatment. CONCLUSIONS: TACE under fluoroscopic guidance in standing horses provided a safe, minimally invasive, and effective method for ICA occlusion and should be recommended for individuals at risk of general anesthesia. Residual neurologic deficits are a common sequela, but they do not reflect a treatment failure.

A minimally-invasive closed chest myocardial occlusion-reperfusion model in rhesus monkeys (Macaca mulatta): monitoring by contrast-enhanced ultrasound imaging.

Myocardial infarction is frequently developed in canine and porcine models but exceptionally in non-human primates. The aim of this study was to develop a minimally invasive myocardial ischemic/reperfusion model in the monkey intended to be combined with imaging techniques, in particular myocardial contrast echocardiography (MCE). A balloon-tipped catheter was advanced via the femoral artery into the left anterior descending artery (LAD) under fluoroscopic guidance in ten anaesthetized male rhesus monkeys (Macaca mulatta). The balloon was inflated to completely occlude the vessel. Coronary angiography (CA) was performed to control the reality of the LAD occlusion/reperfusion. The ischemia period was followed by 3-6 h of reperfusion. Myocardial perfusion was evaluated during ischemia and at reperfusion by MCE using a novel ultrasound contrast agent (BR38). Occlusion was successfully induced during 18-50 min in nine out of the ten evaluated monkeys. ST segment elevation indicated myocardial ischemia. MCE showed complete transmural arrest of myocardial blood flow during the ischemia period and no persistent microvascular perfusion defects during reperfusion. A minimally invasive closed-chest model was successfully developed for creating myocardial ischemia in the rhesus monkey (Macaca mulatta). This technique could have an important role in mimicking acute coronary syndrome under physiologically and ethically-acceptable conditions. MCE provides non-invasively information on myocardial perfusion status, information not available from CA.