Lipid and Corticosteroid Biomarkers Under the Influence of Bisphosphonates.

Detecting the use of bisphosphonates (BPs) in equine athletes is of interest to regulators and laboratories due to the threat to welfare issues for the potential to provide analgesic effects and manipulating bone structure. The detection of BPs in biological matrices is challenging due to erratic biological elimination and inconsistent analytical recoveries. Therefore, complementary approaches are needed to provide evidence of their misuse in racehorses. BPs have two sub-classes: nitrogenous and non-nitrogenous. This study investigated plasma elimination following administration of one example from each sub-class, together with changes in endogenous eicosanoid and corticosteroids. Zoledronic acid (ZA) and tiludronic acid (TA) were administered by IV infusion to 8 thoroughbred horses with an 11-month washout period between each administration. Sample preparation for quantification of BPs by liquid chromatography-tandem mass spectrometry (LC-MS/MS) utilised a two-step solid phase extraction (SPE) consisting of polymeric reversed-phase followed by weak anion exchange prior to derivatisation using trimethyl orthoacetate. Endogenous biomarkers were analysed after protein precipitation and SPE with polymeric reversed-phase prior to liquid chromatography-high resolution mass spectrometry (LC-HRMS) using data independent acquisition. The LC-MS/MS analysis showed ZA was undetectable after 8 h post-administration while TA was detected up to the final collection point of 28 days post-administration. The LC-HRMS analysis utilised targeted (i.e., prior inclusion list of compounds) approaches to monitor level changes of eicosanoid and corticosteroid biomarkers. Putative biomarkers were identified and now subject to validation for translation into routine sample analysis for improved retrospectivity to detecting BP misuse in equine plasma.

Evaluating the Effects of Solid-Phase Cartridge Chemistry on Extraction of Bisphosphonates and ITPP From Equine Plasma.

Bisphosphonate drugs and myo-inositol trispyrophosphate are of concern to the racing industry and have been listed as prohibited substances in equine sports. The current bisphosphonate plasma screening analysis employed at the Australian Racing Forensic Laboratory involves the use of sequential solid-phase extraction procedures, passing the samples through a mixed mode cartridge, followed by a weak anion exchange cartridge. The eluates collected following the second extraction are then methylated and analysed by liquid chromatography-mass spectrometry. Under these extraction conditions, some bisphosphonates have shown poor recovery. To improve the extraction efficacy, the effects of cartridge chemistry were evaluated. In particular, the weak anion exchange cartridges used for screening were compared to an affinisep AttractSPE polymeric phase cartridge. The effectiveness of each extraction approach was assessed through both a visual comparison of signal to noise in extracted chromatograms and recovery measurements to determine the best approach for routine screening.

Enhanced analysis of equine plasma for the presence of recombinant human erythropoietin - Implementation of an improved workflow.

An improved screening workflow and a robust capillary flow LC-MS confirmatory method for the detection of recombinant human erythropoietin (rHuEPO) has been implemented to increase the sensitivity of rHuEPO detection and to reduce the number of suspect samples committed to confirmatory testing. The influence of repeated dosing of epoetin-ß on the detection window of rHuEPO in equine plasma was assessed using the optimised method. Samples were initially assessed using an economical R&D Human EPO Duo-Set ELISA Development System. Samples indicating a result greater than the batch baseline were analysed using the complementary R&D Human EPO Quantikine IVD ELISA kit. All samples recording an abnormal screening result were subjected to confirmatory analysis. Confirmation of rHuEPO in plasma (≥2.5 ml) in the range of 4-13 mIU/ml (n = 6) was achieved using immunoaffinity enrichment, tryptic digestion, and capillary flow LC-MS/MS. Four horses were administered a single dose of epoetin-ß (10,000 IU) via the subcutaneous and intravenous routes, on two occasions, seven days apart. The excretion profile was rapid with epoetin-ß detection times of 48 to 72 h following each administration, with no appreciable difference observed between the two routes of administration. This workflow has been shown as an effective anti-doping strategy related to rHuEPO misuse and supports the use of out-of-competition testing of horses in the 2 to 3-day period prior to race-day.

Confirmation of ethanol administration in racing greyhounds by LC-MS-MS.

Ethanol is a prohibited substance in professional animal racing as its administration causes physiological effects such as depression of the central nervous system. Regulation of potential doping agents, including those that inhibit performance, is critical to ensure integrity and animal welfare in greyhound racing, but the detection of ethanol is complicated by dietary and/or environmental exposure. In response, a reliable analytical method capable of detecting recent ethanol administration in greyhound urine samples was validated and implemented. Liquid chromatography-tandem mass spectrometry (LC-MS-MS) was used to investigate the variation in urinary ethanol metabolites; ethyl-ß-D glucuronide (EG; γ ¯ EG $$ {\overline{\gamma}}_{\mathrm{EG}} $$ = 1.0 µg/ml, s EG $$ {s}_{\mathrm{EG}} $$ = 3.3 µg/ml) and ethyl sulfate (ES; γ ¯ ES $$ {\overline{\gamma}}_{\mathrm{ES}} $$ = 0.9 µg/ml, s ES $$ {s}_{\mathrm{ES}} $$ = 1.9 µg/ml) levels from a reference population of 202 racing greyhounds. These were compared to urine samples collected following administration of ethanol to one male and one female greyhound. Results were used to establish a threshold within the national rules of greyhound racing: γ ¯ EG $$ {\overline{\gamma}}_{\mathrm{EG}} $$ and γ ¯ ES $$ {\overline{\gamma}}_{\mathrm{ES}} $$ > 20 µg/ml in urine are defensible criteria to confirm ethanol administration to greyhounds. Case studies of competition samples are provided to demonstrate the forensic translation of this work.

Exploring the impact of the COVID-19 environment on nursing delivery of family-centred care in a paediatric hospital.

AIMS AND OBJECTIVES: To understand how the pandemic environment impacted the delivery of FCC of children and families from a nursing perspective in a major tertiary paediatric hospital. BACKGROUND: Family-centred care (FCC) is a well-established framework to promote parental involvement in every aspect of a child's hospitalization, however, rules and restrictions in place during the COVID-19 pandemic affected the ways in which Family-centred Care could be delivered in practice. DESIGN: This is a qualitative exploratory descriptive study to elicit the perspective of paediatric nurses delivering care to children in a hospital during the COVID-19 pandemic in Victoria, Australia. METHODS: Nurses from all subspecialties in a tertiary paediatric hospital were invited to participate in virtual focus groups to discuss their experience of delivering FCC during the COVID-19 pandemic. Focus groups were recorded and transcribed, then analysed using Framework Analysis. RESULTS: Nineteen nurses participated across seven focus groups during June and July 2020. The four themes-Advocating with empathy, Enabling communication, Responding with flexibility, and Balancing competing considerations-and the eight subthemes that were generated, outline how nurses deliver FCC, and how these FCC actions were impacted by the COVID-19 environment and the related hospital restrictions. CONCLUSION: This study documents the experiences, resilience and resourcefulness of paediatric nurses in Australia during the COVID-19 pandemic as well as moving Family-centred Care from a theoretical framework into a practical reality. IMPACT: The findings from this study should inform consideration of the impacts of public health policies during infectious disease outbreaks moving forward. In addition by describing the core actions of Family-centred Care, this study has implications for educational interventions on how to translate FCC theory into practice. No public or patient contribution as this study explored nursing perceptions only.

Intrapartum Maternal Cardiac Arrest: A Simulation Case for Multidisciplinary Providers.

Introduction: Cardiac arrest in pregnancy is rare. Advanced Cardiovascular Life Support courses rarely address interventions specific to obstetric patients, and knowledge gaps are frequent among providers. The Society for Obstetric Anesthesia and Perinatology and American Heart Association have published guidelines regarding management of cardiac arrest in pregnancy, and interdisciplinary simulation training has been advocated to reinforce key management points for this clinical scenario. Methods: In situ multidisciplinary simulation training was implemented for anesthesia and maternal fetal medicine fellows and obstetric nurses at our hospital. The case was amniotic fluid embolism in a 35-year-old parturient at term. The patient had a witnessed seizure before cardiovascular collapse. Learners were expected to initiate high-quality cardiopulmonary resuscitation and perform a perimortem cesarean delivery within 5 minutes while demonstrating clear communication with each other. The case required a labor room, high-fidelity mannequin, defibrillator, code cart, cesarean section instruments, and simulated medications and intravenous fluids. Results: Participants comprised two obstetric anesthesia fellows, three maternal fetal medicine fellows, and three obstetric nurses. Positive feedback about the training and increased perceptions of self-efficacy were received. Potential systems issues were detected and corrected because of the training, highlighting the value of in situ drills. Discussion: We found it challenging to implement more frequent multidisciplinary sessions, but participants found the experience highly rewarding. We hope to expand the training to all physicians and nurses covering the unit on a regular basis. Modified scenario versions are being used for nursing-only and obstetric resident-only simulations during protected teaching time for those services.

Detection and quantification of dermorphin and selected analogs in equine urine.

BACKGROUND: Dermorphin, a hepta-peptide with potent analgesic properties, is classified as a doping agent in equine racing. Since its discovery, a number of biologically active structural analogs have been synthesized and made commercially available so there is a need for reliable methods of detection. METHODOLOGY/RESULTS: A sensitive detection method was developed for dermorphin and six analogs in equine urine. Peptide enrichment was achieved using weak cation exchange with subsequent separation and detection by nano-UHPLC-MS/MS. Method validation parameters included: specificity, linearity (5-10000 pg/ml), recovery (58-93%), intra and inter-assay repeatability, LOD (5-50 pg/ml) and matrix effects. CONCLUSION: The presented method will facilitate the control of the abuse of dermorphin and selected analogs in equine sports.

The effect of collaboration on obstetric patient safety in three academic facilities.

Interprofessional collaboration is critical to the provision of safe patient care and provider satisfaction. Collaboration is an active process that can help maximize positive patient outcomes. Three academic institutions implemented collaborative processes as part of their perinatal patient safety initiatives based on anecdotal experiences and safety culture surveys that demonstrated positive outcomes. Reliable tools and additional research are needed to measure the extent and impact of collaboration on patient outcomes in perinatal care.