Label-free Proteomics for Discovering Biomarker Candidates for Controlling Krypton Misuse in Castrated Horses (Geldings).

Recent advances in label-free quantitative proteomics may support its application in identifying and monitoring biomarkers for the purpose of doping control in equine sports. In this study, we developed a workflow of label-free quantitative proteomics to propose plasma protein biomarkers in horses after administration with krypton (Kr), a potential erythropoiesis-stimulating agent. Plasma proteomes were profiled by using nanoliquid chromatography-high-resolution mass spectrometry. An in-house mass spectral library consisting of 1121 proteins was compiled using samples collected from geldings (castrated horses) in the administration trial and geldings in training. A data-independent acquisition method was used to quantify an array of plasma proteins across plasma samples from the administration trial. Statistical analyses proposed a profile of 83 biomarker candidates that successfully differentiated Kr-administered samples from control samples, with the ability to detect Kr exposure for up to 13 days (the last sample collected in the administration trial). The model also correctly classified 32 in-training geldings as untreated controls. This is significantly longer than the 1 h detection time of plasma Kr using headspace gas chromatography-tandem mass spectrometry. Bioinformatic analyses enriched biomarker candidates relevant to complement activation and iron metabolism. The upregulation of transferrin receptor protein 1, one of the candidates related to iron metabolism, in plasma after Kr administration was validated by selected reaction monitoring of corresponding peptides. These results have demonstrated label-free quantitative proteomics as a promising approach to propose plasma protein biomarkers to enhance doping control. Data are available via ProteomeXchange with identifier PXD017262.

The morbidity and mortality outcomes of indigenous Australian peoples after isolated coronary artery bypass graft surgery: the influence of geographic remoteness.

BACKGROUND: Though Indigenous Australian peoples reportedly have poorer survival outcome after cardiac surgery, few studies have jointly documented the experience of major morbidity, and considered the influence of patient geographic remoteness. METHODS: From January 1998 to September 2008, major morbidity events and survival were recorded for 2748 consecutive patients undergoing coronary artery bypass graft surgery. Morbidity and survival analyses adjusted for propensity deciles based on patient ethnicity and age, sex, left ventricular ejection fraction, recent myocardial infarction, tobacco smoking, diabetes, renal disease and history of stroke. Sensitivity analyses controlled for the patient accessibility/remoteness index of Australia (ARIA). RESULTS: The 297 Indigenous Australian patients (10.8% of total) had greater odds for total morbidity (adjusted odds ratio = 1.55; 95% confidence interval [CI] 1.04-2.30) and prolonged ventilation (adjusted odds ratio = 2.08; 95% confidence interval [CI] 1.25-3.44) in analyses adjusted for propensity deciles and geographic remoteness. With a median follow-up of 7.5 years (interquartile range 5.2-10.2), Indigenous Australian patients were found to experience 30% greater mortality risk (unadjusted hazard ratio = 1.30; 95% CI: 1.03-1.64, p = 0.03). The effect size strengthened after adjustment for propensity score (adjusted hazard ratio = 1.49; 95% CI: 1.13-1.96, p = .004). Adjustment for ARIA categorisation strengthened the effect size (adjusted HR = 1.54 (95% CI: 1.11-2.13, p = .009). CONCLUSION: Indigenous Australian peoples were at greater risk for prolonged ventilation and combined morbidity outcome, and experienced poorer survival in the longer term. Higher mortality risk among Indigenous Australians was evident even after controlling for remoteness and accessibility to services.

Generalised urticaria and angioedema complicating switch back from subcutaneous to intravenous vedolizumab therapy.

We report a unique case of a 34-year-old man with ulcerative colitis, previously in complete remission with intravenous vedolizumab monotherapy, who developed an urticarial injection-site reaction on switching to a subcutaneous preparation and thereafter experienced a new hypersensitivity reaction on switch back to intravenous vedolizumab, necessitating complete discontinuation from this drug. This case highlights the need for vigilance on switching back to intravenous preparations of vedolizumab, in response to injection-site reactions with a subcutaneous preparation, even if the intravenous preparation had been previously well tolerated by the patient.

Metabolic studies of selective androgen receptor modulators RAD140 and S-23 in horses.

This paper describes the studies of the in vitro biotransformation of two selective androgen receptor modulators (SARMs), namely, RAD140 and S-23, and the in vivo metabolism of RAD140 in horses using ultra-high performance liquid chromatography-high resolution mass spectrometry. in vitro metabolic studies of RAD140 and S-23 were performed using homogenised horse liver. The more prominent in vitro biotransformation pathways for RAD140 included hydrolysis, hydroxylation, glucuronidation and sulfation. Metabolic pathways for S-23 were similar to those for other arylpropionamide-based SARMs. The administration study of RAD140 was carried out using three retired thoroughbred geldings. RAD140 and the majority of the identified in vitro metabolites were detected in post-administration urine samples. For controlling the misuse of RAD140 in horses, RAD140 and its metabolite in sulfate form gave the longest detection time in hydrolysed urine and could be detected for up to 6 days post-administration. In plasma, RAD140 itself gave the longest detection time of up to 13 days. Apart from RAD140 glucuronide, the metabolites of RAD140 described herein have never been reported before.

Label-free proteomics for discovering biomarker candidates of RAD140 administration to castrated horses.

Selective androgen receptor (AR) modulators (SARMs) are potent anabolic agents with a high potential of misuse in horseracing and equestrian sports. In this study, we applied label-free proteomics to discover plasma protein biomarkers in geldings (castrated horses) after administration with a popular SARM named RAD140. Tryptic peptides were prepared from plasma samples and analyzed by nano-flow ultrahigh-performance liquid chromatography-high-resolution tandem mass spectrometry (nano-UHPLC-HRMS/MS) using data-independent acquisition (DIA) method. Orthogonal projection on latent structure-discriminant analysis (OPLS-DA) has led to the development of a predictive model that could discriminate RAD140-administered samples from control samples and could also correctly classify 18 out of 19 in-training horses as control samples. The model comprises 75 proteins with variable importance in projection (VIP) score above 1. Gene Ontology (GO) enrichment analysis and literature review have identified upregulation of AR-regulated clusterin, and proteins associated with inflammation (haptoglobin, cluster of differentiation 14 [CD14], and inter-alpha-trypsin inhibitor heavy chain 4 [ITIH4]) and erythropoiesis (glycosylphosphatidylinositol-specific phospholipase D1 [GPLD1]) after RAD140 administration. Their changes were confirmed by selected reaction monitoring (SRM) experiments. Similar effects have been reported by the use of androgens and other SARMs. This is the first reported study that describes the use of a proteomic biomarker approach to detect horses that have been administered with RAD140 by applying label-free proteomic profiling of plasma samples. These results support the concept of a biomarker-driven approach to enhance the doping control of RAD140 and potentially other SARMs in the future.

A Rare Case of Severe Nontropical Isolated Right Ventricular Endomyocardial Fibrosis.

We present a case of late presentation nontropical endomyocardial fibrosis isolated to the right ventricle and tricuspid valve (TV). In response to deteriorating hemodynamics, surgical debulking and TV removal were performed before initiation of centralized venoarterial extracorporeal membrane oxygenation support. Definitive endomyocardial resection with a TV prosthesis was then successfully completed. (Level of Difficulty: Advanced.).

First-in-Human Chronic Implant Experience of the Substernal Extravascular Implantable Cardioverter-Defibrillator.

OBJECTIVES: The aim of this study was to evaluate the safety and performance of an extravascular (EV) implantable cardioverter-defibrillator (ICD). BACKGROUND: Limitations of existing transvenous and subcutaneous ICD systems include lead reliability and morbidity issues associated with ICD lead implantation in the vasculature or lack of pacing therapies (e.g., antitachycardia pacing) in subcutaneous systems. The EV defibrillator uses a novel substernal lead placement to address these limitations. METHODS: This was a prospective, nonrandomized, chronic pilot study conducted at 4 centers in Australia and New Zealand. Participants were 21 patients referred for ICD implantation. Patients received EV ICD systems. Data collection included major systemic and procedural adverse events, defibrillation testing at implantation, and sensing and pacing thresholds. RESULTS: Among 20 patients who underwent successful implantation, the median defibrillation threshold was 15 J, and 90% passed defibrillation testing with a ≥10-J safety margin. Mean R-wave amplitude was 3.4 ± 2.0 mV, mean ventricular fibrillation amplitude was 2.8 ± 1.7 mV, and pacing was successful in 95% at ≤10 V. There were no intraprocedural complications. Two patients have undergone elective chronic system removal since hospital discharge. In the 15 patients presently implanted, the systems are stable in long-term follow-up. CONCLUSIONS: This first-in-human evaluation of an EV ICD demonstrated the feasibility of substernal lead placement, defibrillation, and pacing with a chronically implanted system. There were no acute major complications, and pacing, defibrillation, and sensing performance at implantation were successful in most patients. (Extravascular ICD Pilot Study [EV ICD]; NCT03608670).

Detection of bioactive peptides including gonadotrophin-releasing factors (GnRHs) in horse urine using ultra-high performance liquid chromatography-high resolution mass spectrometry (UHPLC/HRMS).

The use of bioactive peptides as a doping agent in both human and animal sports has become increasingly popular in recent years. As such, methods to control the misuse of bioactive peptides in equine sports have received attention. This paper describes a sensitive accurate mass method for the detection of 40 bioactive peptides and two non-peptide growth hormone secretagogues (< 2 kDa) at low pg/mL levels in horse urine using ultra-high performance liquid chromatography-high resolution mass spectrometry (UHPLC/HRMS). A simple mixed-mode cation exchange solid-phase extraction (SPE) cartridge was employed for the extraction of 42 targets and/or their in vitro metabolites from horse urine. The final extract was analyzed using UHPLC/HRMS in positive electrospray ionization (ESI) mode under both full scan and data independent acquisition (DIA, for MS2 ). The estimated limits of detection (LoD) for most of the targets could reach down to 10 pg/mL in horse urine. This method was validated for qualitative detection purposes. The validation data, including method specificity, method sensitivity, extraction recovery, method precision, and matrix effect were reported. A thorough in vitro study was also performed on four gonadotrophin-releasing factors (GnRHs), namely leuprorelin, buserelin, goserelin, and nafarelin, using the S9 fraction isolated from horse liver. The identified in vitro metabolites have been incorporated into the method for controlling the misuse of GnRHs. The applicability of this method was demonstrated by the identification of leuprorelin and one of its metabolites, Leu M4, in urine obtained after intramuscular administration of leuprorelin to a thoroughbred gelding (castrated horse).

Genome Diversity and the Origin of the Arabian Horse.

The Arabian horse, one of the world's oldest breeds of any domesticated animal, is characterized by natural beauty, graceful movement, athletic endurance, and, as a result of its development in the arid Middle East, the ability to thrive in a hot, dry environment. Here we studied 378 Arabian horses from 12 countries using equine single nucleotide polymorphism (SNP) arrays and whole-genome re-sequencing to examine hypotheses about genomic diversity, population structure, and the relationship of the Arabian to other horse breeds. We identified a high degree of genetic variation and complex ancestry in Arabian horses from the Middle East region. Also, contrary to popular belief, we could detect no significant genomic contribution of the Arabian breed to the Thoroughbred racehorse, including Y chromosome ancestry. However, we found strong evidence for recent interbreeding of Thoroughbreds with Arabians used for flat-racing competitions. Genetic signatures suggestive of selective sweeps across the Arabian breed contain candidate genes for combating oxidative damage during exercise, and within the "Straight Egyptian" subgroup, for facial morphology. Overall, our data support an origin of the Arabian horse in the Middle East, no evidence for reduced global genetic diversity across the breed, and unique genetic adaptations for both physiology and conformation.

Vettath's technique of long mammary patch reconstruction of a diffusely diseased left anterior descending coronary artery without endarterectomy on the beating heart.

Surgery for diffuse coronary artery disease requires coronary reconstruction with or without endarterectomy. Considering the immediate and late postoperative problems of coronary endarterectomy, many surgeons now prefer coronary reconstruction without endarterectomy. Patch reconstruction of the diseased coronary artery with either the internal mammary artery or another conduit is an attractive option. This article describes the technique of long mammary patch reconstruction of the left anterior descending coronary artery without endarterectomy as an off-pump procedure.

Tuberculosis of thymus--a case report.

Tuberculosis rarely figures in the list of thymic masses. We hereby report a case of thymic tuberculosis successfully managed with surgical excision and antituberculouschemotherapy.

Use of extracorporeal life support for emergency coronary artery bypass grafting.

A 14-year old boy was admitted with an anomalous origin of the right coronary artery (RCA) from the left sinus of Valsalva, with an interarterial course of the narrow proximal segment. He underwent coronary ostial augmentation and main pulmonary artery translocation to the left pulmonary artery. In the post-cardiopulmonary bypass (CPB) period, he developed thrombotic occlusion of the RCA resulting in arrhythmia and ventricular dysfunction, requiring extracorporeal life support (ECLS) in the form of extracorporeal membrane oxygenation rather than CPB. After confirming this complication by cardiac catheterization, the right coronary artery territory was revascularized with a pedicled right internal thoracic artery graft. The entire procedure was performed on a beating heart using a stabilizer during ECLS. This strategy may be useful in situations with unstable haemodynamics and a low risk of blood loss.

Extracorporeal life support in the acute management of tumour lysis syndrome.

A 16-month old boy presented with a severe tumour lysis syndrome (TLS) complicating induction therapy for acute myeloid leukaemia. This was further complicated by a respiratory syncytial virus infection. The failure of response to escalating treatment necessitated the use of extracorporeal life support (ECLS) during continuation of his induction chemotherapy. He was weaned from support after the resolution of the TLS and completed chemotherapy successfully. ECLS may have a role to play in the acute support of children with cardiorespiratory failure as a result of malignancy or the required treatment.

Mammary artery patch reconstruction of left anterior descending coronary artery.

Most patients with diffuse coronary disease require endarterectomy for revascularization. In view of the high morbidity and mortality associated with coronary endarterectomy, patch angioplasty and reconstruction of the coronary artery was developed. We retrospectively evaluated 104 patients who underwent mammary artery patch angioplasty of the left anterior descending coronary artery. The procedure consisted of laying open the entire diseased segment of the coronary artery and patching it with an in-situ left internal mammary artery onlay patch, without endarterectomy, in an off-pump procedure. One (0.9%) patient died, 2 (1.8%) suffered perioperative myocardial infarction, and an intraaortic balloon pump was used in 2. The mean blood loss was 224 mL. Intensive care unit stay was 2.5 days, and hospital stay was 7.8 days. At 1-year follow-up, most patients were in New York Heart Association functional class I. Follow-up angiography was carried out in 16 patients and showed good patency of all anastomoses. Arterial patch reconstruction of the left anterior descending coronary artery can be performed safely and effectively in an off-pump procedure, with excellent early and intermediate results.

Simultaneous off-pump coronary artery bypass grafting and ascending aortobifemoral bypass graft via ventral abdominal route.

Aortoiliac disease often coexists with coronary artery disease. It is not uncommon to subject a patient to two separate interventions. We report two cases in which in simultaneous off-pump coronary artery bypass grafting was done with an ascending aortobifemoral bypass graft through the ventral abdominal route without any additional morbidity. Combining a technically simple method of limb bypass with an off-pump cardiac surgery is a promising procedure for revascularization of myocardium and lower limbs. We discuss the merits of combining an off-pump coronary artery bypass grafting procedure with a limb bypass.

Traumatic suprahepatic inferior vena cava injury: surgical management.

SUMMARY: : Suprahepatic inferior vena caval (IVC) injuries are rare but carry nearly a 100% mortality rate. The main problem with its surgical management is the technical difficulty in draining the IVC during cardiopulmonary bypass. In this report, an efficient method of IVC drainage for repair of the IVC on cardiopulmonary bypass is described.