Doping control of estra-4,9-diene-3,17-dione in horses.

Estra-4,9-diene-3,17-dione (dienedione) is an anabolic-androgenic steroid (AAS) available on the market as a dietary supplement for bodybuilding. It is prohibited in both human and equine sports due to its potential performance-enhancing effect. With the rare presence of the 4,9-diene configuration in endogenous steroids, dienedione has been considered as a synthetic AAS. Nevertheless, the reoccurring detection of dienedione in entire male horse urine samples led to the investigation of its possible endogenous nature in horses, and its endogenous nature in entire male horses has been recently confirmed and reported by the authors' laboratory. While dienedione is not detected in castrated horses (geldings), it is essential to study its elimination and identify its metabolites for its effective control. To study the elimination and biotransformation of dienedione, administration experiments were performed by giving three castrated horses (geldings) each single oral dose of 1500 mg of dienedione powder for seven consecutive days. The postulated in vivo metabolites included 17-hydroxyestra-4,9-dien-3-one (M1a and M1b), hydroxylated dienedione (M2a, M2b, M3a, M3b, M4, M5) and hydroxylated M1 (M6a, M6b, M7a, M7b, M8a and M8b), formed from hydroxylation and reduction of dienedione. To control the misuse of dienedione in geldings, M3a and M3b are the potential targets that gave the longest detection time, which could be detected for up to 2-5 days in urine and 0.4-4 days in plasma.

The Feasibility and Impact of Practising Online Forest Bathing to Improve Anxiety, Rumination, Social Connection and Long-COVID Symptoms: A Pilot Study.

BACKGROUND: Long-COVID affects over 144 million people globally. In the absence of treatments, there is a need to establish the efficacy of therapies that improve patient outcomes. Forest bathing has been demonstrated to improve physical and mental outcomes but there is no evidence in Long-COVID patients. Accordingly, this pilot study sought to determine the feasibility and effectiveness of online forest bathing in adults with Long-COVID. METHODS: Feasibility was assessed by monitoring retention rates and participant feedback. In a waitlist controlled, repeated measures design, 22 Long-COVID patients completed weekly online surveys during a four-week waitlist control period, before engaging in four weekly online forest bathing sessions, completing post-intervention surveys following each session. RESULTS: In terms of retention, 27% did not provide post-intervention data, reasons for non-adherence were: feeling too ill, having medical appointments, or having career responsibilities. Compared with the waitlist control period, there were statistically significant improvements in Anxiety (49% decrease), Rumination (48% decrease), Social Connection (78% increase), and Long-COVID symptoms (22% decrease). Written qualitative comments indicated that participants experienced feelings of calm and joy, felt more connected socially and with nature, and experienced a break from the pain and rumination surrounding their illness. CONCLUSIONS: Online Forest bathing resulted in significant improvements in well-being and symptom severity and could be considered an accessible and inexpensive adjunct therapy for Long-COVID patients. Where people have limited access to in-person nature, virtual nature may offer an alternative to improve health and well-being outcomes.

Optimization and implementation of four duplex quantitative polymerase chain reaction assays for gene doping control in horseracing.

The concern about gene doping has remained high in horseracing and other equestrian competitions. Our laboratory has previously developed a duplex quantitative polymerase chain reaction (qPCR) assay capable of detecting in equine blood the human erythropoietin (hEPO) transgene and equine tubulin α 4a (TUBA4A) gene as an internal control the latter providing quality control over DNA extraction and qPCR. This study aimed to optimize the method for routine testing of regulatory samples. The use of an automated DNA extraction system has increased the sample throughput, consistency of DNA extraction, and recovery of reference materials. The use of reduced concentration of primers and hydrolysis probe for internal control minimized their competition with transgene amplification and improved the assay sensitivity. Spike-in of an exogenous internal control at low concentration for plasma analysis has also been validated. Using the new workflow, four duplex qPCR assays have been developed for the detection of transgenes, namely, hEPO, human growth hormone (hGH), insulin-like growth factor 1 (hIGF-1), and equine EPO (eEPO). The estimated limits of detection (LODs) of each transgene were 2000 copies/mL of blood and 200 copies/mL of plasma. This method could detect the presence of transgene in blood and plasma collected from a horse administered intramuscularly (IM) with recombinant adeno-associated virus (rAAV) carrying the hEPO transgene. A longer detection time was observed in blood than in plasma. The methods have been applied to the screening of over a thousand official racehorse samples since June 2020 for the presence of these transgenes.

Comparison of FreeHand® robot-assisted with human-assisted laparoscopic fundoplication.

OBJECTIVE: To compare the safety, operative time and feasibilty of FreeHand® robot (FreeHand Ltd, Guildford, United Kingdom) with manual camera control approach for Laparoscopic Fundoplication. MATERIAL AND METHODS: A case control study was performed for patients undergoing laparoscopic fundoplication. Primary outcome was operative time; secondary outcomes included length of stay, post-operative morbidity, symptoms at first follow-up and total post-operative out-patient visits. RESULTS: Forty-four patients underwent laparoscopic fundoplication between January 2014 and June 2016. Twenty-six (59%) underwent conventional human-assisted fundoplication while 18 (41%) had FreeHand® robot assisted procedures. Mean operative time for conventional laparoscopic fundoplication was 165 min compared with 129 min in the robot-assisted group, saving 36 min (p < .001).The median length of stay was 1.5 days in the robot-assisted as compared to two days in the conventional group. Sixteen percent of robot-assisted as opposed to 30% of conventional group patients experienced complications. There was no 30-day mortality. Two patients required more than one post-operative clinic visit in robot-assisted against six in conventional group. CONCLUSION: Robot-assisted fundoplication is safe, feasible and reduces operative time. Furthermore, this negates need of assistant. Mean operative time for robot-assisted fundoplication was 36 min less than for conventional fundoplication. Advantages also include fewer adverse events, shorter length of stay and less post-operative clinic visits.

The Role of Empathy in Professional Quality of Life: a Study on Australian Police Officers Working in Sexual Assault and Child Abuse Investigation.

Police working in sexual assault and child abuse investigation may be at risk of secondary trauma effects and burnout, particularly if they do not have protective mechanisms in place. Empathy has shown to be vital in protecting against secondary trauma and burnout, as well as enhancing compassion satisfaction. The current cross-sectional study surveyed 216 Australian police participants working in sexual assault and child abuse investigation exploring the relationship between different facets of empathy and professional quality of life factors. All facets of empathy predicted compassion satisfaction and negatively predicted burnout. Aspects of cognitive empathy negatively predicted burnout and secondary traumatic stress, while aspects of emotional/physiological and cognitive empathy positively predicted compassion satisfaction. Novel gender differences were found, with males at higher risk of burnout; tenure significantly predicted burnout and secondary traumatic stress; and results supported that empathy is a vital mechanism for sustaining wellbeing, satisfaction, and efficacy in this work.

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.

Super-Obesity in the Elderly: Is Bariatric Surgery Justified?

Although the prevalence of obese elderly patients is increasing, the outcomes of bariatric surgery in this potentially high-risk cohort remain poorly understood, especially those relating to quality of life. Furthermore, there is no data on the efficacy of bariatric surgery in the super-obese elderly. We identified 50 consecutive patients undergoing bariatric surgery aged 60 years or over, and compared the outcomes of the super-obese (BMI ≥ 50; n = 26) with those of BMI < 50. Mean follow-up was 33 months. There were no significant differences between the groups in terms of comorbidities, operation-type and peri-operative complications. Mean percentage excess weight loss was comparable between the groups (56.7 vs. 58.8 %; p = 0.81), as was resolution of comorbidities and post-operative quality of life (mean Bariatric Analysis and Reporting Outcome System (BAROS) 3.5 vs. 3.1; p = 0.64).

Microwave processing of gustatory tissues for immunohistochemistry.

We use immunohistochemistry to study taste cell structure and function as a means to elucidate how taste receptor cells communicate with nerve fibers and adjacent taste cells. This conventional method, however, is time consuming. In the present study we used taste buds from rat circumvallate papillae to compare conventional immunohistochemical tissue processing with microwave processing for the colocalization of several biochemical pathway markers (PLCß2, syntaxin-1, IP3R3, α-gustducin) and the nuclear stain, Sytox. The results of our study indicate that in microwave versus conventional immunocytochemistry: (1) fixation quality is improved; (2) the amount of time necessary for processing tissue is decreased; (3) antigen retrieval is no longer needed; (4) image quality is superior. In sum, microwave tissue processing of gustatory tissues is faster and superior to conventional immunohistochemical tissue processing for many applications.

Intra-abdominal collections following laparoscopic versus open appendicectomy: an experience of 516 consecutive cases at a district general hospital.

BACKGROUND: The objective of this study is to evaluate the incidence of intra-abdominal collections (IACs) in all patients undergoing laparoscopic (LA) and open appendicectomy (OA) from April 2009 to October 2011 in a district general hospital with expertise in minimally invasive surgery (MIS). METHODS: A retrospective review of all patients undergoing appendicectomy in the specified time period was carried out. IACs were identified from various in-hospital data resources. Severity of appendicitis was assessed from histology reports. RESULTS: 516 patients were identified, of whom 242 (47 %) underwent OA and 274 (53 %) LA. Twenty-six (5 %) patients were found to have IACs postoperatively. Fifteen (5.5 %) IACs were identified in the laparoscopic group and 11 (4.5 %) in the open group. There was no statistically significant difference in the risk of developing IACs in open versus laparoscopic groups [odds ratio (OR) 1.22, confidence interval (CI) 0.55-2.70, P = 0.63]. Patients were twelve times more likely to develop IACs with an appendix identified as being necrotic or perforated on histology (OR 12.24, CI 5.29-28.32, P < 0.0001). There was a trend towards shorter total hospital stay in the LA (3.58 days, CI 3.0-4.1 days) compared with OA (4.31 days, CI 3.7-4.9 days, P = 0.082) group, although this was not statistically significant. CONCLUSIONS: Increased rates of IAC following LA have been identified in some studies. Our series shows that, in a centre with adequate MIS experience, the IAC rate following LA is comparable to that of the open approach and should not deter surgeons with adequate support and resources.

Immunocytochemical analysis of P2X2 in rat circumvallate taste buds.

BACKGROUND: Our laboratory has shown that classical synapses and synaptic proteins are associated with Type III cells. Yet it is generally accepted that Type II cells transduce bitter, sweet and umami stimuli. No classical synapses, however, have been found associated with Type II cells. Recent studies indicate that the ionotropic purinergic receptors P2X2/P2X3 are present in rodent taste buds. Taste nerve processes express the ionotropic purinergic receptors (P2X2/P2X3). P2X2/P2X3(Dbl-/-) mice are not responsive to sweet, umami and bitter stimuli, and it has been proposed that ATP acts as a neurotransmitter in taste buds. The goal of the present study is to learn more about the nature of purinergic contacts in rat circumvallate taste buds by examining immunoreactivity to antisera directed against the purinergic receptor P2X2. RESULTS: P2X2-like immunoreactivity is present in intragemmal nerve processes in rat circumvallate taste buds. Intense immunoreactivity can also be seen in the subgemmal nerve plexuses located below the basal lamina. The P2X2 immunoreactive nerve processes also display syntaxin-1-LIR. The immunoreactive nerves are in close contact with the IP(3)R3-LIR Type II cells and syntaxin-1-LIR and/or 5-HT-LIR Type III cells. Taste cell synapses are observed only from Type III taste cells onto P2X2-LIR nerve processes. Unusually large, "atypical" mitochondria in the Type II taste cells are found only at close appositions with P2X2-LIR nerve processes. P2X2 immunogold particles are concentrated at the membranes of nerve processes at close appositions with taste cells. CONCLUSIONS: Based on our immunofluorescence and immunoelectron microscopical studies we believe that both perigemmal and most all intragemmal nerve processes display P2X2-LIR. Moreover, colloidal gold immunoelectron microscopy indicates that P2X2-LIR in nerve processes is concentrated at sites of close apposition with Type II cells. This supports the hypothesis that ATP may be a key neurotransmitter in taste transduction and that Type II cells release ATP, activating P2X2 receptors in nerve processes.

An audit of post-splenectomy prophylaxis--are we following the guidelines?

INTRODUCTION: Asplenic individuals have major difficulties coping with specific infections (e.g. Streptococcus pneumoniae). This is an audit to look at a district general hospital's compliance with published guidelines for immunisations and antibiotic prophylaxis post-splenectomy. PATIENTS AND METHODS: A retrospective review of hospital records of consecutive splenectomy patients from January 1996 to March 2001. RESULTS: Of 76 patients, 72% were vaccinated (30/76 with pneumococcal, HIB and meningococcal vaccines, 15/76 with Pneumovax and HIB, 10/76 with Pneumovax only), 63% were discharged on prophylactic antibiotics, and 81% of surviving patients had adequate communication with the GP regarding splenectomy. Patients undergoing non-elective splenectomy were less likely to be vaccinated or receive prophylactic antibiotics when compared with elective splenectomy patients. CONCLUSIONS: Results are comparable with other published studies, but are still unsatisfactory for many splenectomy patients. Vaccination rates must be improved and more information given to patients and GPs to allow for appropriate follow-up care.