Reggie-1/flotillin-2 promotes secretion of the long-range signalling forms of Wingless and Hedgehog in Drosophila

The lipid-modified morphogens Wnt and Hedgehog diffuse poorly in isolation yet can spread over long distances in vivo, predicting existence of two distinct forms of these mophogens. The first is poorly mobile and activates short-range target genes. The second is specifically packed for efficient spreading to induce long-range targets. Subcellular mechanisms involved in the discriminative secretion of these two forms remain elusive. Wnt and Hedgehog can associate with membrane microdomains, but the function of this association was unknown. Here we show that a major protein component of membrane microdomains, reggie-1/flotillin-2, plays important roles in secretion and spreading of Wnt and Hedgehog in Drosophila. Reggie-1 loss-of-function results in reduced spreading of the morphogens, while its overexpression stimulates secretion of Wnt and Hedgehog and expands their diffusion. The resulting changes in the morphogen gradients differently affect the short- and long-range targets. In its action reggie-1 appears specific for Wnt and Hedgehog. These data suggest that reggie-1 is an important component of the Wnt and Hedgehog secretion pathway dedicated to formation of the mobile pool of these morphogens

Peri-operative red blood cell transfusion in neonates and infants: NEonate and Children audiT of Anaesthesia pRactice IN Europe: A prospective European multicentre observational study

BACKGROUND: Little is known about current clinical practice concerning peri-operative red blood cell transfusion in neonates and small infants. Guidelines suggest transfusions based on haemoglobin thresholds ranging from 8.5 to 12 g dl-1, distinguishing between children from birth to day 7 (week 1), from day 8 to day 14 (week 2) or from day 15 (≥week 3) onwards. OBJECTIVE: To observe peri-operative red blood cell transfusion practice according to guidelines in relation to patient outcome. DESIGN: A multicentre observational study. SETTING: The NEonate-Children sTudy of Anaesthesia pRactice IN Europe (NECTARINE) trial recruited patients up to 60 weeks' postmenstrual age undergoing anaesthesia for surgical or diagnostic procedures from 165 centres in 31 European countries between March 2016 and January 2017. PATIENTS: The data included 5609 patients undergoing 6542 procedures. Inclusion criteria was a peri-operative red blood cell transfusion. MAIN OUTCOME MEASURES: The primary endpoint was the haemoglobin level triggering a transfusion for neonates in week 1, week 2 and week 3. Secondary endpoints were transfusion volumes, 'delta haemoglobin' (preprocedure - transfusion-triggering) and 30-day and 90-day morbidity and mortality. RESULTS: Peri-operative red blood cell transfusions were recorded during 447 procedures (6.9%). The median haemoglobin levels triggering a transfusion were 9.6 [IQR 8.7 to 10.9] g dl-1 for neonates in week 1, 9.6 [7.7 to 10.4] g dl-1 in week 2 and 8.0 [7.3 to 9.0] g dl-1 in week 3. The median transfusion volume was 17.1 [11.1 to 26.4] ml kg-1 with a median delta haemoglobin of 1.8 [0.0 to 3.6] g dl-1. Thirty-day morbidity was 47.8% with an overall mortality of 11.3%. CONCLUSIONS: Results indicate lower transfusion-triggering haemoglobin thresholds in clinical practice than suggested by current guidelines. The high morbidity and mortality of this NECTARINE sub-cohort calls for investigative action and evidence-based guidelines addressing peri-operative red blood cell transfusions strategies. TRIAL REGISTRATION: ClinicalTrials.gov, identifier: NCT02350348

Successful resuscitation and neurological monitoring of a dog with out-of-hospital cardiopulmonary arrest due to pentobarbital overdose.

OBJECTIVE To describe the clinical signs, electroencephalographic (EEG) findings, treatment, and outcome in a dog after successful resuscitation from out-of-hospital cardiopulmonary arrest (OHCA) induced by pentobarbital intoxication. CASE SUMMARY A 10-year-old, male intact Jack Russell Terrier was referred for management of refractory status epilepticus and presented dead on arrival. After 7 minutes of cardiopulmonary resuscitation, return of spontaneous circulation was achieved, but the dog remained comatose, apneic, and lacked brainstem reflexes on neurological examination 6 hours following resuscitation. Magnetic resonance imaging showed polioencephalomalacia consistent with prolonged epileptiform activity, and EEG was initially concerning for electrocerebral inactivity. Following supportive care that included short-term mechanical ventilation, the dog made a full recovery and was discharged from the hospital alive 7 days postresuscitation. It was later revealed that the dog had been administered an unknown amount of pentobarbital during transportation, which likely contributed to the OHCA, clinical, and EEG findings. NEW INFORMATION PROVIDED This is the first report to describe the full recovery and hospital discharge of a dog suffering OHCA and the first description of EEG findings in a clinical veterinary patient following cardiopulmonary arrest and successful resuscitation. Factors likely contributing to successful patient outcome and potential benefits and limitations of EEG in monitoring postcardiac arrest patients are discussed

Successful resuscitation and neurological monitoring of a dog with out-of-hospital cardiopulmonary arrest due to pentobarbital overdose.

ObjectiveTo describe the clinical signs, electroencephalographic (EEG) findings, treatment, and outcome in a dog after successful resuscitation from out-of-hospital cardiopulmonary arrest (OHCA) induced by pentobarbital intoxication.Case summaryA 10-year-old, male intact Jack Russell Terrier was referred for management of refractory status epilepticus and presented dead on arrival. After 7 minutes of cardiopulmonary resuscitation, return of spontaneous circulation was achieved, but the dog remained comatose, apneic, and lacked brainstem reflexes on neurological examination 6 hours following resuscitation. Magnetic resonance imaging showed polioencephalomalacia consistent with prolonged epileptiform activity, and EEG was initially concerning for electrocerebral inactivity. Following supportive care that included short-term mechanical ventilation, the dog made a full recovery and was discharged from the hospital alive 7 days postresuscitation. It was later revealed that the dog had been administered an unknown amount of pentobarbital during transportation, which likely contributed to the OHCA, clinical, and EEG findings.New information providedThis is the first report to describe the full recovery and hospital discharge of a dog suffering OHCA and the first description of EEG findings in a clinical veterinary patient following cardiopulmonary arrest and successful resuscitation. Factors likely contributing to successful patient outcome and potential benefits and limitations of EEG in monitoring postcardiac arrest patients are discussed

Musculoskeletal Load in and Highly Repetitive Actions of Animal Facility Washroom Employees

Regular work tasks in the washroom of laboratory animal facilities include cleaning of cages and bottles and handling of chow and bedding. These operations largely are carried out by hand. We quantitatively determined the musculoskeletal load on the trunk and upper limbs of washroom employees in an animal facility with a holding capacity of 35,000 rodent cages by using a computer-assisted, quantitative, recording, and long-term analysis (CUELA) system, which volunteers wore during routine work. Parallel video recording allowed exact assignment of each movement of body and limbs to the data recorded by the sensors. For the most part, trunk movements were unassociated with risk of injury. Evaluation of upper limb movements by CUELA indicated elevated burden on shoulder, elbows, and wrists due to the high repetitiveness and range of movements and postures. However, after additional work factors like low effort and the presence of micropauses were taken into account, workers were not at risk for the development of musculoskeletal disorders of the upper limbs. Handling bottles, chow, and bedding and maneuvering trolleys that entailed greater musculoskeletal loads did not yield evidence of overstraining, because the actions typically were executed alternately and were of short duration during daily shifts. The results represent quantitative information on the musculoskeletal load of regular washroom operations in a laboratory animal facility. These data provide the basis for ergonomic redesign of operations and implementation of automation for highly repetitive movements

Self-reported clinical practice of small animal cardiopulmonary resuscitation and compliance with RECOVER guidelines among veterinarians in eight Western European regions

Introduction: The objective of this study was to assess whether small animal veterinarians across Western Europe are compliant with the 2012 cardiopulmonary resuscitation (CPR) guidelines by the Reassessment Campaign on Veterinary Resuscitation (RECOVER). Methods: A previously published online questionnaire from Switzerland was adapted and translated into 7 languages, corresponding to national languages in Austria, France, Germany, Ireland, Italy, Liechtenstein, Netherlands, Portugal, Spain, and the United Kingdom. The survey was distributed via respective national veterinary organizations and social media outlets. A subset of questions was analyzed to evaluate respondent demographics, RECOVER guideline awareness, and to allocate composite compliance scores for CPR preparedness, basic life support (BLS) and advanced life support (ALS). Percentages of group total (95% confidence interval) were calculated. Multivariable logistic regression was used to evaluate the effects of region of practice, gender, age, specialty training, and guideline awareness on compliance. Odds ratios (95% confidence interval) were generated and significance set at P < 0.05. Results: Nine-hundred and thirty respondents were included in analysis. Awareness of and compliance with RECOVER guidelines varied widely across regions. Compliance with all assessed RECOVER guideline recommendations was highest in Germany/Austria [14% (7- 27%)] and lowest in France and Portugal [0% (0–3%)]. CPR preparedness compliance was higher in participants aware of RECOVER guidelines [OR 10.1 (5.2-19.5)], those practicing in Germany/Austria [OR 4.1 (1.9–8.8)] or UK/Ireland [OR 2.2 (1.3–3.7)], and lower in those practicing in Portugal [OR 0.2 (0.1–0.9)]. Specialty training [OR 1.8 (1.1–2.9)], guideline awareness [OR 5.2 (3.2–8.6)], and practice in Germany/Austria [OR 3.1 (1.5–6.5)], UK/Ireland [OR 2.6 (1.7–4.1)], or the Netherlands [OR 5.3 (2.0–14.2)] were associated with increased BLS compliance. ALS compliance was higher in participants with guideline awareness [OR 7.0 (2.9–17.0)], specialty training [OR 6.8 (3.8–12.1)], those practicing in Germany/Austria [OR 3.5 (1.3–9.6)], UK/Ireland [OR 4.0 (1.9–8.3)], or Spain [OR 3.2 (1.2–8.3)] and in younger survey participants [OR 0.9 (0.9–1.0)]. Conclusions: Awareness and compliance with RECOVER guidelines varied widely among countries surveyed, however overall compliance scores in all countries were considered low. Further research may highlight factors surrounding poor guideline awareness and compliance so targeted efforts can be made to improve veterinary CPR in Europe

Magnetic resonance T2 mapping and diffusion-weighted imaging for early detection of cystogenesis and response to therapy in a mouse model of polycystic kidney disease

Polycystic kidney disease (PKD) is among the leading causes of end-stage renal disease. Increasing evidence exists that molecular therapeutic strategies targeted to cyst formation and growth might be more efficacious in early disease stages, highlighting the growing need for sensitive biomarkers. Here we apply quantitative magnetic resonance imaging techniques of T2 mapping and diffusion-weighted imaging in the jck mouse model for PKD using a clinical 3.0 T scanner. We tested whether kidney T2 values and the apparent diffusion coefficient (ADC) are superior to anatomical imaging parameters in the detection of early cystogenesis, as shown on macro- and histopathology. We also tested whether kidney T2 values and ADC have the potential to monitor early treatment effects of therapy with the V2 receptor antagonist Mozavaptane. Kidney T2 values and to a lesser degree ADC were found to be highly sensitive markers of early cystogenesis and superior to anatomical-based imaging parameters. Furthermore, kidney T2 values exhibited a nearly perfect correlation to the histological cystic index, allowing a clear separation of the two mouse genotypes. Additionally, kidney T2 values and ADC were able to monitor early treatment effects in the jck mouse model in a proof-of-principle experiment. Thus, given the superiority of kidney T2 values and ADC over anatomical-based imaging in mice, further studies are needed to evaluate the translational impact of these techniques in patients with PKD