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Background: Torus (buckle) fractures of the wrist are the most common fractures in children, involving the distal radius and/or ulna. It is unclear if children require rigid immobilisation and follow-up, or would equally recover by being discharged without any immobilisation or a bandage. Given the large number of these injuries, identifying the optimal treatment strategy could have important effects on the child, school absences and NHS costs. Objectives: To establish whether treating children with a distal radius torus fracture with the offer of a soft bandage and immediate discharge (“offer of bandage”), provides the same recovery, in terms of pain, function, complications, acceptability, school absence and resource use, as treatment with rigid immobilisation and follow-up as per usual practice (“rigid immobilisation”). Design: A pragmatic, multicentre, randomised controlled equivalence trial. Setting: 23 UK Emergency Departments. Participants: 965 children (4-15 years) with a distal radius torus fracture were randomised from January 2019 to July 2020 using a secure, centralised, online encrypted randomisation service. Exclusion criteria included presentation >36 hours after injury, multiple injuries and inability to complete follow-up. Interventions: Offer of bandage (n= 489) of whom the bandage was applied in 458, versus rigid immobilisation (n= 476). Participants and clinicians were not blinded to the treatment allocation. Main outcome measures: Pain at 3-days post-randomisation measured using the WongBaker FACES Pain Rating Scale. Secondary outcomes were the PROMIS Upper Extremity score for children, health related quality of life (QoL), complications, school absence, analgesia use and resource use collected up to 6 weeks post-randomisation. Results: 94% of participants provided primary outcome data. Pain was equivalent at the 3 day primary outcome for both groups. With reference to the pre-specified equivalence margin of 1.0, the adjusted difference in the intention-to-treat population was -0.10; 95% CI -0.37 to 0.17 and the per-protocol population was -0.06; 95% CI -0.34 to 0.21. There was equivalence in pain for both age subgroups (4-7 and 8–15 years). There was no difference in the rate of complications, with five (1.0%) in the offer of bandage group and three (0.6%) in the rigid immobilisation group. There were no differences between treatment groups in functional recovery, QoL or school absence at any point during follow-up. Analgesia use was marginally higher at Day 1 in the offer of bandage group (83% vs 78% participants), though was no different at other times. The offer of bandage significantly reduced the cost of treatment, and had a high probability of cost-effectiveness. Limitations: Families had a strong pre-existing preference for rigid immobilisation. Given this, and the inability to blind families to the treatment allocation, observer bias was a concern. However, there was clear evidence of equivalence. Conclusions: The study findings support the offer of bandage in children with a distal radius torus fracture.

Osteoarthritis (OA) is a chronic joint disease caused by both mechanical damage and metabolic factors, which intertwine in their pathogenetic pathways. We hypothesise that the oxidised cholesterol derivative, 7-ketocholesterol (7-KCh), represents a danger signal in the synovia of patients with OA and promotes low-grade inflammation. The study would aim to elucidate the possible immune mechanisms initiated by 7-KCh that contribute to oxidative stress and inflammation in the synovia and synovial CD68+ cells, which are mostly macrophages. The polarisation of synovial CD68+ cells, their tissue distribution in relation to T and natural killer (NK) cells, and the influence of 7-KCh on the phenotype and intracellular cytokine and chemokine production in suspension is worth analysing. We envisage that this research would contribute to a better understanding of the biology of CD68+ macrophages influenced by 7-KCh and encourage the development of therapeutic approaches based on directing macrophage polarisation.

Spondyloarthritis (SpA) is a group of autoinflammatory/autoimmune diseases that affects the spine and peripheral joints. Involvement of the cervical spine correlates with occurrence of temporomandibular disorders (TMDs). Proinflammatory cytokines, interleukin (IL)-23, IL-17A, and tumour necrosis factor-alpha (TNF-α) are significantly involved in the immunopathogenesis of SpA, and inflammation is one of the causes of TMDs. We hypothesised that internal derangement (ID) of temporomandibular joints (TMJs), as the most frequent TMDs in patients with SpA, could be associated with salivary pro-inflammatory markers and clinical activity of axial SpA, measured with laboratory methods or rheumatological indexes. If a statistically significant difference in the concentration of salivary IL-23, IL-17A or TNF-α exists between patients with symptomatic or asymptomatic ID and patients without TMDs, the diagnostic accuracy of certain cytokine(s) for ID can be calculated. The diagnostic accuracy of the cytokine(s) could emphasise the inflammatory background of ID and possibly broaden therapeutic approaches for symptomatic ID in patients with SpA with biological drugs in addition to the routine application of occlusal splint and/or physical therapy.

To compare characteristics of temporomandibular disorders (TMDs) in patients with rheumatoid arthritis (RA) to controls without RA.The sample included 80 subjects (aged 33 to 73 years; 88% women), 40 in each group. An international diagnostic protocol for TMD was followed.Arthralgia was the most prevalent TMD in the RA group. Orofacial pain was more common than in controls (42.5% vs 15%, P = .031), with higher chronic pain grade and pain intensity (P ≤ .005). Somatization and depression were also increased (P.001). In multiple logistic regression analysis, arthralgia (OR: 6.4; 95% CI: 1.1 to 37.1; P = .038) and age ≥ 55 years were predictors of RA (OR 3.9; 95% CI: 1.4 to 10.8; P = .009) when controlling for the effects of gender and pain intensity. TMD was related to 7.4-times higher odds for presence of orofacial pain, while RA was related to 3.4-times higher odds for pain.RA patients experienced more orofacial pain and higher pain intensity, somatization, and depression compared to healthy individuals. Pain is more influenced by TMD than by RA.

Aim: Investigations have shown that for the antibody–drug conjugate (ADC) belantamab mafodotin, concentrations of the cysteine-conjugated metabolite, Cys-mcMMAF, were overestimated in the presence of the ADC during sample processing when utilizing a historical SPE method. Results: A new assay was developed utilizing an acidic protein precipitation to remove the ADC early in the extraction process, thus eliminating the risk of overestimating Cys-mcMMAF in the presence of belantamab mafodotin. In vitro experiments demonstrated a linear relationship between the concentration of belantamab mafodotin and the release of Cys-mcMMAF. Extensive stability assessments were performed to cover storage of study samples. Conclusion: This work emphasized the critical importance of understanding the performance of a bioanalytical method for free toxic payload in the presence of the ADC.

Purpose Sotrovimab (VIR-7831), a human IgG1κ monoclonal antibody (mAb), binds to a conserved epitope on the SARS-CoV-2 spike protein receptor binding domain (RBD). The Fc region of VIR-7831 contains an LS modification to promote neonatal Fc receptor (FcRn)–mediated recycling and extend its serum half-life. Here, we aimed to evaluate the impact of the LS modification on tissue biodistribution, by comparing VIR-7831 to its non-LS-modified equivalent, VIR-7831-WT, in cynomolgus monkeys. Methods 89Zr-based PET/CT imaging of VIR-7831 and VIR-7831-WT was performed up to 14 days post injection. All major organs were analyzed for absolute concentration as well as tissue:blood ratios, with the focus on the respiratory tract, and a physiologically based pharmacokinetics (PBPK) model was used to evaluate the tissue biodistribution kinetics. Radiomics features were also extracted from the PET images and SUV values. Results SUVmean uptake in the pulmonary bronchi for 89Zr-VIR-7831 was statistically higher than for 89Zr-VIR-7831-WT at days 6 (3.43 ± 0.55 and 2.59 ± 0.38, respectively) and 10 (2.66 ± 0.32 and 2.15 ± 0.18, respectively), while the reverse was observed in the liver at days 6 (5.14 ± 0.80 and 8.63 ± 0.89, respectively), 10 (4.52 ± 0.59 and 7.73 ± 0.66, respectively), and 14 (4.95 ± 0.65 and 7.94 ± 0.54, respectively). Though the calculated terminal half-life was 21.3 ± 3.0 days for VIR-7831 and 16.5 ± 1.1 days for VIR-7831-WT, no consistent differences were observed in the tissue:blood ratios between the antibodies except in the liver. While the lung:blood SUVmean uptake ratio for both mAbs was 0.25 on day 3, the PBPK model predicted the total lung tissue and the interstitial space to serum ratio to be 0.31 and 0.55, respectively. Radiomics analysis showed VIR-7831 had mean-centralized PET SUV distribution in the lung and liver, indicating more uniform uptake than VIR-7831-WT. Conclusion The half-life extended VIR-7831 remained in circulation longer than VIR-7831-WT, consistent with enhanced FcRn binding, while the tissue:blood concentration ratios in most tissues for both drugs remained statistically indistinguishable throughout the course of the experiment. In the bronchiolar region, a higher concentration of 89Zr-VIR-7831 was detected. The data also allow unparalleled insight into tissue distribution and elimination kinetics of mAbs that can guide future biologic drug discovery efforts, while the residualizing nature of the 89Zr label sheds light on the sites of antibody catabolism.

Background The effect of frailty and poor cardiovascular health on mortality for males and females is not fully elucidated. We investigated whether the combined burden of frailty and poor cardiovascular health is associated with all-cause and cardiovascular disease (CVD) mortality by sex and age. Methods We analyzed data of 35,207 non-institutionalized US residents aged 20–85 years old (mean age [standard deviation]: 46.6 [16.7 years], 51.4% female, 70.8% White, 10.3% Black, 13.2% Hispanic) from the National Health and Nutrition Examination Survey (1999–2015). Cardiovascular health was measured with the American Heart Association’s Life’s Simple 7 score (LS7). A 33-item frailty index (FI) was constructed to exclude cardiovascular health deficits. We grouped the FI into 0.1 increments (non-frail: FI < 0.10, very mildly frail: 0.1 ≤ FI < 0.20, mildly frail: 0.20 ≤ FI < 0.30, and moderately/severely frail: FI ≥ 0.30) and LS7 into tertiles (T1[poor] = 0–7, T2[intermediate] = 8-9, T3[ideal] = 10–14). All-cause and CVD mortality data were analyzed up to 16 years. All regression models were stratified by sex. Results The average FI was 0.09 (SD 0.10); 29.6% were at least very mildly frail, and the average LS7 was 7.9 (2.3). Mortality from all-causes and CVD were 8.5% (4228/35,207) and 6.1% (2917/35,207), respectively. The median length of follow-up was 8.1 years. The combined burden of frailty and poor cardiovascular health on mortality risk varied according to age in males (FI*age interaction p = 0.01; LS7*age interaction p < 0.001) but not in females. In females, poor FI and LS7 combined to predict all-cause and CVD mortality in a dose-response manner. All-cause and CVD mortality risk was greater for older males (60 and 70 years old) who were at least mildly frail and had intermediate cardiovascular health or worse (hazard ratio [lower/higher confidence interval ranges] range: all-cause mortality = 2.02–5.30 [1.20–4.04, 3.15–6.94]; CVD-related mortality = 2.22–7.16 [1.03–4.46, 4.49–11.50]) but not for younger males (30, 40, and 50 years old). Conclusions The combined burden of frailty and LS7 on mortality is similar across all ages in females. In males, this burden is greater among older people. Adding frailty to assessments of overall cardiovascular health may identify more individuals at risk for mortality and better inform decisions to implement preventative or treatment approaches.

We aimed to evaluate the diagnostic accuracy of the proinflammatory monocyte chemotactic protein-1 (MCP-1) in the diagnosis of asymptomatic diastolic dysfunction (DD) in patients with psoriatic arthritis (PsA). The disease activity in psoriatic arthritis (DAPSA) was determined using clinical and laboratory parameters, and echocardiography was performed to estimate DD. Serum MCP-1 concentrations were elevated in PsA patients with DD diagnosed with ultrasound (median (25th percentile, 75th percentile): 366.6 pg/mL (283, 407.1 pg/mL) vs. 277.5 pg/mL (223.5, 319.1 pg/mL) in controls; P < 0.0017 ). PsA patients with serum MCP-1 concentration higher than the cut-off value of 347.6 pg/mL had a 7.74-fold higher chance of developing DD than PsA patients with lower serum MCP-1 concentrations (controls), with a specificity of 86.36% and sensitivity of 55%, as verified using ultrasound. The group with MCP-1 concentrations above the cut-off value also showed a higher late peak diastolic mitral inflow velocity, A-wave value ( P = 0.000005 ), E/E ′ ratio ( P = 0.00005 ), and a lower E/A ratio ( P = 0.000002 ), peak systolic left atrial reservoir strain, SA value ( P = 0.0066 ), early peak diastolic displacement of the mitral septal annulus, E ′ wave value ( P = 0.003 ), than controls. Systolic blood pressure ( P = 0.01 ), LDL cholesterol concentration ( P = 0.012 ), glucose concentration ( P = 0.011 ), and DAPSA ( P = 0.0000 ) increased in the PsA group with higher MCP-1 concentrations, although there were no differences in comorbidities and therapy between the groups compared. Thus, the serum MCP-1 concentration was a significant and independent prognostic indicator for asymptomatic DD in PsA patients ( area under the curve = 0.730 , P = 0.001 ). The DAPSA score in PsA patients might indicate the need for echocardiography and adjustment of anti-inflammatory treatment in terms of DD prevention.

Background physical activity reduces frailty in community-dwelling older adults. How exercise influences frailty in hospitalised older adults requires additional investigation. Objectives (i) to examine the impact of an exercise intervention on frailty in older adults admitted to an acute care ward, and (ii) to determine the impact of baseline frailty on the effectiveness of this intervention. Setting/participants this is a secondary analysis of a randomised controlled clinical trial that tested an intensive exercise intervention in ≥75-year-old adults admitted to an acute care ward. Methods the intervention included two daily sessions of moderate-intensity exercises (control received usual care). A 63-item Frailty Index (FI) was constructed, and three groups were formed: Results a total of 323 individuals were included. The mean age was 87.1 years (± 4.8 standard deviation [SD]) and 56.3% were females. The intervention group improved FI from 0.26 (± 0.10 SD) to 0.20 (± 0.10 SD), whereas the control group FI worsened from 0.25 (± 0.1 SD) to 0.27 (± 0.10 SD). After stratifying by baseline FI, SPPB and depression improved in the intervention group across all levels of frailty; FI, BI and quality of life only improved in individuals with a baseline FI ≥ 0.2. Conclusions frailty improves with an intensive individualised exercise intervention, especially in those with high baseline levels of frailty. In addition, frailty is a useful outcome when examining the impact of an intervention of hospitalised older adults.

Aim: Recent advances in microflow ultra performance liquid chromatography (UPLC) systems offer higher sensitivity with robustness to meet the routine bioanalytical demands. Modern high-resolution mass spectrometers (HRMS) enable the development of highly selective methods with broad dynamic range. Results: The quantitative performances of tandem quadrupole MS and HRMS were comprehensively compared using seven intact peptide hormones up to 9.4 kDa. Results show comparable performance between two platforms in sensitivity, accuracy and linearity. For some peptides, HRMS provided lower background interference. The benefit of increased sensitivity using microflow UPLC was also demonstrated. Conclusion: HRMS is a versatile platform capable of both basic characterization and reliable quantitation in complex matrices. Microflow UPLC provides lower LLOQs than conventional flow systems, even with less sample volume injected.

Objective The associations of moderate‐vigorous physical activity (MVPA) bouts and patterns of sedentary time (ST) with frailty according to cardiovascular disease (CVD) status are unknown. Methods Accelerometry in adults ≥50 years old from the 2003‐2004 and 2005‐2006 National Health and Nutrition Examination Survey were used. Bouted and sporadic MVPA in ≥10‐minute or 0.05 for CVD interaction). Frequency of ST breaks was not associated with frailty. Average ST break intensity was protective in both groups. The duration of breaks in ST was associated with lower frailty in CVD participants only (P > 0.05 for CVD interaction). Conclusion Insufficient MVPA and prolonged ST are detrimental despite CVD status.

IntroductionIn the past 20 years, the increasing burden of heart disease in an ageing population has resulted in cardiac surgery (CS) being offered to more frail and older patients with multiple comorbidities. Frailty and malnutrition are key geriatric syndromes that impact postoperative outcomes, including morbidity, mortality and prolonged hospital length of stay. Enhanced recovery protocols (ERPs), such as prehabilitation, have been associated with a reduction in complications after CS in vulnerable patients. The use of nutritional ERPs may enhance short-term and long-term recovery and mitigate frailty progression while improving patient-reported outcomes.Methods and analysisThis trial is a two-centre, double-blinded, placebo, randomised controlled trial with blinded endpoint assessment and intention-to-treat analysis. One-hundred and fifty CS patients will be randomised to receive either a leucine-rich protein supplement or a placebo with no supplemented protein. Patients will consume their assigned supplement two times per day for approximately 2 weeks pre-procedure, during in-hospital postoperative recovery and for 8 weeks following discharge. The primary outcome will be the Short Physical Performance Battery score. Data collection will occur at four time points including baseline, in-hospital (pre-discharge), 2-month and 6-month time points post-surgery.Ethics and disseminationThe University of Manitoba Biomedical Research Ethics Board (20 March 2018) and the St Boniface Hospital Research Review Committee (28 June 2019) approved the trial protocol for the primary site in Winnipeg, Manitoba, Canada. The second site’s (Montreal, Quebec) ethics has been submitted and pending approval from the Research Ethics and New Technology Development Committee for the Montreal Heart Institute (December 2020). Recruitment for the primary site started February 2020 and the second site will begin January 2021. Data gathered from the PROTein to Enhance outComes of (pre)frail paTients undergoing Cardiac Surgery Study will be published in peer-reviewed journals and presented at national and international conferences. Knowledge translation strategies will be created to share findings with stakeholders who are positioned to implement evidence-informed change.Potential study impactMalnutrition and frailty play a crucial role in post-CS recovery. Nutritional ERPs are increasingly being recognised as a clinically relevant aspect of perioperative care. As such, this trial is to determine if leucine-rich protein supplementation at key intervals can mitigate frailty progression and facilitate enhanced postoperative recovery.Trial registration numberClinicalTrials.gov Registry (NCT04038294).

The main risk factor for primary open-angle glaucoma (POAG) is increased intraocular pressure (IOP). It is of interest that about half of the patients have an IOP within the normal range (normal-tension glaucoma, NTG). Additionally, there is a group of patients with a high IOP but no glaucomatous neurodegeneration (ocular hypertension, OHT). Therefore, risk factors other than IOP are involved in the pathogenesis of glaucoma. Since the retina has a very high oxygen-demand, decreased autoregulation and a fluctuating oxygen supply to the retina have been linked to glaucomatous neurodegeneration. To assess the significance of these mechanisms, we have utilized a human experimental model, in which we stress participants with a fluctuating oxygen supply. Levels of oxidative stress molecules, antioxidants, and lipid mediators were measured in the plasma. Patients with NTG, OHT, and control subjects were found to have similar levels of oxidative stress markers. In contrast, patients with OHT had a higher level of total antioxidant capacity (TAC) and pro-homeostatic lipid mediators. Thus, we suggest that OHT patients manage fluctuating oxygen levels more efficiently and, thus, are less susceptible to glaucomatous neurodegenerations, due to enhanced systemic antioxidant protection.

Costa, EC, Kent, DE, Boreskie, KF, Hay, JL, Kehler, DS, Edye-Mazowita, A, Nugent, K, Papadopoulos, J, Stammers, AN, Oldfield, C, Arora, RC, Browne, RAV, and Duhamel, TA. Acute effect of high-intensity interval versus moderate-intensity continuous exercise on blood pressure and arterial compliance in middle-aged and older hypertensive women with increased arterial stiffness. J Strength Cond Res 34(5): 1307-1316, 2020-Hypertension and arterial stiffness are common in middle-aged and older women. This study compared the acute effect of high-intensity interval exercise (HIIE) and moderate-intensity continuous exercise (MICE) on blood pressure (BP) and arterial compliance in middle-aged and older hypertensive women with increased arterial stiffness. Nineteen women (67.6 ± 4.7 years) participated in this randomized controlled crossover trial. Subjects completed a control, MICE (30 minutes at 50-55% of heart rate reserve [HRR]), and HIIE (10 × 1 minute at 80-85% of HRR, 2 minutes at 40-45% of HRR) session in random order. Blood pressure and large and small arterial compliance (radial artery pulse wave analysis) were measured at baseline and 30, 60, 90, and 120 minutes after sessions. A p0.05 was considered statistically significant. Systolic BP was reduced in ∼10 mm Hg after MICE at 30 minutes and after HIIE at all time points (30, 60, 90, and 120 minutes) after exercise compared with the control session (p0.05). Only HIIE showed lower systolic BP levels at 60, 90, and 120 minutes after exercise compared with the control session (∼10 mm Hg; p0.05). No changes were observed in diastolic BP, or in large and small arterial compliance (p0.05). High-intensity interval exercise elicited a longer systolic postexercise hypotension than MICE compared with the control condition, despite the absence of acute modifications in large and small arterial compliance.

Background: Physical activity is important in the management of frailty. Here we determine if specific frailty deficits relate to lower activity across degrees of frailty, and what types of physical activities are commonly reported. Methods: Accelerometer data from 6817 adults 20–85 years old (2003–2004/2005–2006 cycles) of the National Health and Nutrition Examination Survey were analyzed. Moderate-to-vigorous intensity physical activity (MVPA) was measured in 1-minute bouts. Frailty was measured with a 46-item frailty index consisting of chronic conditions, healthcare utilization, difficulties in activities of daily living (ADL), and laboratory deficits. Individuals were stratified into frailty groups: non-frail, 0.20 (21%; n = 1431). Linear regression models adjusted for age, sex, demographics, and accelerometer wear time. Results: Among the total sample, the presence of individual deficits were generally associated with lower MVPA. The presence of more chronic conditions, ADLs, abnormal laboratory values, and greater healthcare utilization had an independent dose association with lower MVPA. The effect of frailty deficits on MVPA were attenuated when examining individuals by frailty level. The number of ADLs, but not the number of chronic conditions or number of laboratory deficits, were independently associated with a lower MVPA level across frailty levels. Healthcare use was associated with less MVPA in the lower frailty groups. The two most popular reported physical activities were walking and cycling across all frailty levels. Conclusions: Deficits in ADLs, but not chronic conditions or laboratory deficits, were consistently associated with lower MVPA across frailty levels.

ObjectivesThe psychological impact of the COVID-19 pandemic on doctors is a significant concern. Due to the emergence of multiple pandemic waves, longitudinal data on the impact of COVID-19 are vital to ensure an adequate psychological care response. The primary aim was to assess the prevalence and degree of psychological distress and trauma in frontline doctors during the acceleration, peak and deceleration of the COVID-19 first wave. Personal and professional factors associated with psychological distress are also reported.DesignA prospective online three-part longitudinal survey.SettingAcute hospitals in the UK and Ireland.ParticipantsFrontline doctors working in emergency medicine, anaesthetics and intensive care medicine during the first wave of the COVID-19 pandemic in March 2020.Primary outcome measuresPsychological distress and trauma measured using the General Health Questionnaire-12 and the Impact of Events-Revised.ResultsThe initial acceleration survey distributed across networks generated a sample of 5440 doctors. Peak and deceleration response rates from the original sample were 71.6% (n=3896) and 56.6% (n=3079), respectively. Prevalence of psychological distress was 44.7% (n=1334) during the acceleration, 36.9% (n=1098) at peak and 31.5% (n=918) at the deceleration phase. The prevalence of trauma was 23.7% (n=647) at peak and 17.7% (n=484) at deceleration. The prevalence of probable post-traumatic stress disorder was 12.6% (n=343) at peak and 10.1% (n=276) at deceleration. Worry of family infection due to clinical work was the factor most strongly associated with both distress (R2=0.06) and trauma (R2=0.10).ConclusionFindings reflect a pattern of elevated distress at acceleration and peak, with some natural recovery. It is essential that policymakers seek to prevent future adverse effects through (a) provision of vital equipment to mitigate physical and psychological harm, (b) increased awareness and recognition of signs of psychological distress and (c) the development of clear pathways to effective psychological care.Trial registration numberISRCTN10666798.

ZusammenfassungFür Diagnostik, Differenzialdiagnostik und Differenzialtherapie des Normaldruckhydrozephalus ist die Bildgebung entscheidend. Auch wenn die Therapie eines Normaldruckhydrozephalus nicht erfolgreich sein sollte, werden zahlreiche Fragen an den Radiologen gestellt. Der Artikel gibt eine Übersicht über das Krankheitsbild des Normaldruckhydrozephalus und die speziellen prä- und postchirurgischen Anforderungen an die Bildgebung.

ObjectivesTo determine the need for recovery (NFR) among emergency physicians and to identify demographic and occupational characteristics associated with higher NFR scores.DesignCross-sectional electronic survey.SettingEmergency departments (EDs) (n=112) in the UK and Ireland.ParticipantsEmergency physicians, defined as any registered physician working principally within the ED, responding between June and July 2019.Main outcome measureNFR Scale, an 11-item self-administered questionnaire that assesses how work demands affect intershift recovery.ResultsThe median NFR Score for all 4247 eligible, consented participants with a valid NFR Score was 70.0 (95% CI: 65.5 to 74.5), with an IQR of 45.5–90.0. A linear regression model indicated statistically significant associations between gender, health conditions, type of ED, clinical grade, access to annual and study leave, and time spent working out-of-hours. Groups including male physicians, consultants, general practitioners (GPs) within the ED, those working in paediatric EDs and those with no long-term health condition or disability had a lower NFR Score. After adjusting for these characteristics, the NFR Score increased by 3.7 (95% CI: 0.3 to 7.1) and 6.43 (95% CI: 2.0 to 10.8) for those with difficulty accessing annual and study leave, respectively. Increased percentage of out-of-hours work increased NFR Score almost linearly: 26%–50% out-of-hours work=5.7 (95% CI: 3.1 to 8.4); 51%–75% out-of-hours work=10.3 (95% CI: 7.6 to 13.0); 76%–100% out-of-hours work=14.5 (95% CI: 11.0 to 17.9).ConclusionHigher NFR scores were observed among emergency physicians than reported in any other profession or population to date. While out-of-hours working is unavoidable, the linear relationship observed suggests that any reduction may result in NFR improvement. Evidence-based strategies to improve well-being such as proportional out-of-hours working and improved access to annual and study leave should be carefully considered and implemented where feasible.

The toxic hallucinogen 25B-NBOMe is very rapidly degraded by human liver microsomes and has low oral bioavailability. Herein we report on the synthesis, microsomal stability, and 5-HT2A/5-HT2C receptor profile of novel analogues of 25B-NBOMe modified at the primary site of metabolism. Although microsomal stability could be increased while maintaining potent 5-HT2 receptor agonist properties, all analogues had an intrinsic clearance above 1.3 L/kg/h predictive of high first-pass metabolism.

Background: FTIH studies can be challenging due to the varying dosing regimens and rapid data delivery. Chemists are asked to provide ultra-low limits of quantitation to provide an understanding of patient efficacy and safety in order to progress drug development. In a recent dermal study it became necessary to reduce the LLOQ of a small molecule drug from 50 to 1 pg/ml due to reductions in the dose and surface area of drug application. Methodology: The 50-fold increase in assay sensitivity necessitated the use of a high-resolution mass spectrometer (LC–HRMS) to separate matrix interferences observed when using a unit resolution triple quadrupole MS. Conclusion: A sensitive, robust assay was validated to support of a FTIH study using a LC–HRMS.

Background: For quantitative bioanalysis utilizing MS, the instrument of choice is typically a triple quadruple mass spectrometer. However, advances in high-resolution MS have allowed sensitivity and dynamic ranges to approach that of triple quadrupole instruments. Results: A matrix-free protein digest, a digested therapeutic protein and the intact peptide therapeutic liraglutide were each analyzed on high-resolution and triple quadrupole mass spectrometers with data compared. Samples from a mouse PK study with liraglutide were analyzed using the two different instruments, and equivalent PK exposure data were demonstrated. Conclusion: High-resolution and triple quadrupole mass spectrometers can generate data resulting in identical PK parameters from an in-life sample set, thus giving confidence in either technique in support of biotherapeutic PK exposure studies.

The neighbourhood environment may enable active aging by allowing the integration of walking into an older adult's daily routine. This study explores the relationship between the neighbourhood built environment and walking among a small group of older adults in a large suburban municipality in Canada. In-depth interviews using a photo-voice approach revealed that the participants walked largely to accumulated physical activity. Older adults who lived in either conventional residential or condominium neighbourhoods discussed poor traffic conditions and lack of benches/trees/places as barriers, and proximity to parks and access to shops as enablers to walking. Poor sidewalk quality, absence of street lights and personal safety concerns were major barriers to walking only for those living in suburban residential neighbourhoods. Our results indicate that high quality- and safe walking infrastructure may facilitate walking for physical activity among older adults living in the suburban communities.

Engaging in health-promoting behaviours has health benefits for people with prediabetes or diabetes. People experience negative affect after diagnoses, which can impede self-regulation of health behaviours. Self-compassion, extending care to oneself in difficult times, can mitigate negative affect and promote self-regulation. This scoping review explored the relationship between self-compassion and adaptive affect, self-regulation and engagement in health-promoting/management behaviours among people with prediabetes or diabetes. We conducted a scoping literature search from 6 databases for studies and conference abstracts. Randomized controlled trials and cross-sectional, longitudinal, observational and qualitative designs focused on self-compassion were included. Eligible studies included adults with diabetes (prediabetes, type 1, type 2 and gestational), measured self-compassion using a validated Self-Compassion Scale (quantitative) or included the 3 components (qualitative) and investigated: negative affect, health promoting/management behaviours and/or self-regulation. After deduplication, 5,338 quantitative and 953 qualitative abstracts, and 18 conference proceedings were screened leaving 35 articles. Full-text screening retained 11 eligible studies (6 cross-sectional studies, 2 randomized controlled trials, 2 longitudinal studies and 1 qualitative study). Higher self-compassion was associated with decreased negative affect in 9 studies and was positively associated with well-being in 1 cross-sectional study. Self-compassion led to decreased negative affect and improved blood glucose in 2 interventions. Five studies found positive associations between self-compassion and health-promoting/management behaviours. One qualitative study found self-compassion to benefit affective reactions, health-promoting behaviours and self-regulation. This review shows that self-compassion is linked to adaptive behavioural and affective responding among people with prediabetes and diabetes, and the need for more research on self-compassion and self-regulation in these populations.

BackgroundPoint Of Care Ultrasound (POCUS) is a potentially useful addition to the technical armamentarium of PEM clinicians. Evidence is accumulating on diagnostic and procedural applications; however despite widespread use in other countries, usage patterns are not clear in our setting. Given the lack of a standardised PEM POCUS curriculum, best practice standards, and formal training courses, we aimed to determine variability in PEM POCUS practices across our PEM research network.Method and resultsThis online survey was distributed in March 2019, with content derived iteratively by the study team from existing literature and input from PERUKI members. One response was sought from each site, to describe department practices, hardware, and major enablers and obstacles. Results are presented using descriptive statistics.Conclusions59/63 (94%) sites responded, including a mix of site and department types (eg 40% tertiary hospital, 60% mixed adult/paediatric EDs, 30% major trauma centres). Almost all (90%) had access to POCUS, and 70% reported limited use by a small proportion of staff, with no named lead. Most (95%) did not have a teaching program or identified trainer. Approximately half (55%) routinely documented findings, most often in clinical notes, and most (65%) did not store images; most (68%) had no quality assurance measures. The greatest enablers of PEM POCUS were its need, funding for equipment, and clinician engagement; the greatest obstacles were availability of training courses and trainers, and time to learn and maintain skills due to service delivery needs.Whilst hardware is available in most EDs, PEM POCUS is not yet widespread, with training being the greatest obstacle. Future work exploring clinician opinions will inform whether the specialty wishes to embrace POCUS, and training course content. However, if adopted, it must be coupled with implementation of best practice standards in domains including quality assurance and governance.

Acute myocardial infarction (AMI) occurs as a result of insufficient myocardial perfusion leading to cell necrosis. This is most commonly due to the obstruction of the coronary artery by ruptured atherosclerotic plaque and thrombosis. Damaged ischemic and necrotic myocardial cells release pro- inflammatory substances in tissue and plasma, leading to a systemic inflammatory response. Profound systemic inflammatory response during ischemia/reperfusion injury causes disruption of endothelial glycocalyx and detachment of endothelial cells that express von Willebrant factor (vWF). We hypothesize that circulating vWF+ endothelial cells could act as antigen presenting cells which interact with T and NK cells directly, by cell to cell contact and indirectly by cytokine and chemokine secretion, leading to the immune response towards inflammation. Analyzing the frequency, phenotype and pro- inflammatory substances produced in circulating vWF positive (+) cells in patients with AMI could be beneficial to determine the severity of the pro- inflammatory response, according to the level of endothelial dysfunction in the early period of AMI. To evaluate these hypotheses, we suggest to determine frequency, phenotype, and ability of cytokine/chemokine production in circulating vWF+ endothelial cells by simultaneous surface and intracellular cell staining, and flow cytometry analysis. Secretion of pro- inflammatory cytokines and chemokines, pro- atherogenic substances and the components of glycocalyx might be measured in supernatants of magnetically separated or sorted vWF+ endothelial cells, as well as in the serum of a patient with acute AMI by enzyme linked– immunoassay tests. The interaction of increasing concentrations of isolated circulating vWF+ endothelial cells and cognate T and NK cells might be investigated by lymphocyte proliferation rate, cytotoxic mediators’ expression, and cytokine production. If our hypothesis is correct, characterization of circulating vWF+ endothelial cells could grant us greater insight into their role in pathophysiology of AMI and the degree of myocardial damage.

Summary of commentary Perioperative neuromuscular electrical stimulation did not avoid postoperative muscle wasting or preserve strength to a greater extent than control patients. Future investigation is needed to determine if neuromuscular electrical stimulation is efficacious in frail patients. However, preoperative exercise training may be needed to preserve skeletal muscle and improve prognosis.

Aim: Typically, quantitation of biotherapeutics from biological matrices by LC–MS is based on a surrogate peptide approach to determine molecule concentration. Recent efforts have focused on quantitation of the intact protein molecules or larger mass subunits of monoclonal antibodies. To date, there has been limited guidance for large or intact protein mass quantitation for quantitative bioanalysis. Methodology: Intact- and subunit-level analyses of biotherapeutics from biological matrices are performed at 12–25 kDa mass range with quantitation data presented. Results: Linearity, bias and other metrics are presented along with recommendations made on the viability of existing quantitation approaches. Conclusion: This communication is intended to start a discussion around intact protein data analysis and processing, recognizing that other published contributions will be required.

IntroductionEfforts to identify individuals at a higher risk for adverse cardiovascular outcomes focus on traditional risk factors, such as age, sex, smoking status, blood pressure and and cholesterol; however, this approach does not directly assess cardiovascular function and may underestimate the risk of experiencing adverse cardiovascular outcomes in women. This prospective, observational cohort study will examine the ability of the Heart Attack Prevention Program for You (HAPPY) Hearts screening protocol, a series of non-invasive procedures to identify middle-aged and older women who are at an elevated risk for experiencing an adverse cardiovascular event in the 5-year period after screening. The predictive value of the HAPPY Hearts protocol will also be compared with the Framingham Risk Score to determine the sensitivity for estimating risk for an adverse cardiovascular outcome.Methods and analysisOne thousand women 55 years of age or older will be recruited to be screened by the HAPPY Hearts protocol. This involves the cardiovascular assessment of resting blood pressure, blood pressure response to 3 min of moderate intensity exercise and large and small arterial elasticity. The participants will be classified into risk categories based on these measures. The incidence of the following adverse cardiovascular outcomes will be assessed in the 5-year period after screening in both groups: ischaemic heart disease, acute myocardial infarction, stroke, percutaneous coronary intervention, coronary bypass surgery, congestive heart failure and new hypertension.Ethics and disseminationInformation gathered in this research will be published in peer-reviewed journals and presented in a programme evaluation report to inform Manitoba Health and key stakeholders about the outcomes of the study. The University of Manitoba Health Research Ethics Board has approved the study protocol V.2.0, dated 29 September 2014 (H2014:224).Trial registration numberNCT02863211.

The sarco(endo)plasmic reticulum calcium ATPase (SERCA) is responsible for transporting calcium (Ca2+) from the cytosol into the lumen of the sarcoplasmic reticulum (SR) following muscular contraction. The Ca2+sequestering activity of SERCA facilitates muscular relaxation in both cardiac and skeletal muscle. There are more than 10 distinct isoforms of SERCA expressed in different tissues. SERCA2a is the primary isoform expressed in cardiac tissue, whereas SERCA1a is the predominant isoform expressed in fast-twitch skeletal muscle. The Ca2+sequestering activity of SERCA is regulated at the level of protein content and is further modified by the endogenous proteins phospholamban (PLN) and sarcolipin (SLN). Additionally, several novel mechanisms, including post-translational modifications and microRNAs (miRNAs) are emerging as integral regulators of Ca2+transport activity. These regulatory mechanisms are clinically relevant, as dysregulated SERCA function has been implicated in the pathology of several disease states, including heart failure. Currently, several clinical trials are underway that utilize novel therapeutic approaches to restore SERCA2a activity in humans. The purpose of this review is to examine the regulatory mechanisms of the SERCA pump, with a particular emphasis on the influence of exercise in preventing the pathological conditions associated with impaired SERCA function.

Objective Cerebrospinal fluid (CSF) overdrainage is a major problem in shunt therapy for hydrocephalus. The adjustable gravitational valve proSA allows for the first time a targeted compensation for overdrainage in the upright position without interfering with the differential pressure valve. To evaluate benefit, safety and reliability, the multicenter prospective registry PROSAIKA was conducted in 10 German neurosurgical centers. Methods Between March 2009 and July 2010, 120 hydrocephalic patients undergoing first time shunt implantation or shunt revision using proSA entered the study. 93 patients completed the 12 months follow-up. Results Hydrocephalus symptoms were improved in 86%, unchanged in 9% and deteriorated in 3%. In 51%, the proSA opening pressure was readjusted one or several times to treat suspected suboptimal shunt function, this resulted in clinical improvement in 55%, no change in 25%, and deterioration in 20% of these patients. The 1 year censored proSA shunt survival rate was 89%. Device related shunt failure was seen in two cases. Conclusions This is the first clinical report on the implantation of the adjustable gravitational valve proSA with a follow-up of 12 months in a substantial number of patients. Irrespective of different hydrocephalus etiologies and indications for shunt surgery, the overall results after 12 months were very satisfying. The high frequency of valve readjustments underlines the fact that preoperative selection of the appropriate valve opening pressure is difficult. The low number of revisions and complications compared to other valves proves that proSA implantation adds no further risk; this valve is reliable, helpful and safe.

The current study evaluated 17 loads of cull beef cows transported in Canadian winter conditions to assess in-transit temperature and humidity, evaluation of events during loading and unloading, and animal condition and bruising. Regardless of the use of boards to block ventilation holes in trailers, temperatures were higher within trailers than at ambient locations during both travel and stationary periods (P < 0.01). Boarding was associated with smaller differences in trailer temperature, compared with ambient conditions, while the trailer was traveling at highway speeds versus when trailers were stationary (P < 0.01). Moisture levels within trailers were not different from ambient conditions when loads using boarding were traveling (P < 0.01), whereas loads without boarding had a larger difference (P < 0.01). The moisture within trailers relative to ambient conditions increased when trailers were stationary compared with traveling when boarding was used (P < 0.01). The majority of cattle transported were in good body condition (97.4% within BCS of 2 to 3.5) and had calm temperaments (96.7%). Although all comparisons were made, only the doghouse compartment had an increased risk of severe bruising compared with all other compartments (odds ratio [95% confidence interval]: 3.0 [1.6–5.5], 3.7 [2.1–6.4], 2.2 [1.3–3.7] and 3.8 [1.5–9.6] in comparison with the back, belly, deck, and nose compartments, respectively; P < 0.05). Increasing the duration of waiting to unload 30 min relative to a 1 h duration increased the odds of severe bruising by 1.18 times (95% confidence interval: 1.09–1.29; P < 0.01). Scoring systems that have been developed for auditing unloading of cattle had limited variation across loads at both loading and unloading. Pretransport assessment of animal condition using the American Meat Institute’s compromised animal score was the only scoring system that was consistent with posttransport scores. We inferred from the temperature and humidity data in the current study that under commercial conditions, boarding may increase ventilation within trailers during travel and decrease ventilation during stationary periods. The current study provides the first indication that issues in Canadian cull cow transport may be related to pretransport animal condition and management of unloading.

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ObjectivesThe objective of this systematic review was to study the impact of preoperative physical activity levels on adult cardiac surgical patients’ postoperative: (1) major adverse cardiac and cerebrovascular events (MACCEs), (2) adverse events within 30 days, (3) hospital length of stay (HLOS), (4) intensive care unit length of stay (ICU LOS), (5) activities of daily living (ADLs), (6) quality of life, (7) cardiac rehabilitation attendance and (8) physical activity behaviour.MethodsA systematic search of MEDLINE, Embase, AgeLine and Cochrane library for cohort studies was conducted.ResultsEleven studies (n=5733 patients) met the inclusion criteria. Only self-reported physical activity tools were used. Few studies used multivariate analyses to compare active versus inactive patients prior to surgery. When comparing patients who were active versus inactive preoperatively, there were mixed findings for MACCE, 30 day adverse events, HLOS and ICU LOS. Of the studies that adjusted for confounding variables, five studies found a protective, independent association between physical activity and MACCE (n=1), 30-day postoperative events (n=2), HLOS (n=1) and ICU LOS (n=1), but two studies found no protective association for 30-day postoperative events (n=1) and postoperative ADLs (n=1). No studies investigated if activity status before surgery impacted quality of life or cardiac rehabilitation attendance postoperatively. Three studies found that active patients prior to surgery were more likely to be inactive postoperatively.ConclusionDue to the mixed findings, the literature does not presently support that self-reported preoperative physical activity behaviour is associated with postoperative cardiac surgical outcomes. Future studies should objectively measure physical activity, clearly define outcomes and adjust for clinically relevant variables.RegistrationTrial registration numberNCT02219815. PROSPERO number CRD42015023606.

Newcastle disease (ND) still remains one of the most important diseases affecting domestic poultry in Colombia. Here, for the first time, we report on the molecular characterization of 12 virulent and 12 avirulent or lentogenic avian paramyxovirus type 1 (APMV-1) strains that were isolated from commercial, backyard, and game poultry in Colombia from 2008 to 2010. The 12 virulent isolates had a fusion (F) protein cleavage site with basic amino acids at positions 113, 115, and 116 and a phenylalanine at position 117 (112RRQKR*F117), characteristic of virulent strains. The remaining 12 isolates had the F protein cleavage sites 112GKQGR*L117 or 112GRQGR*L117 typical of avirulent or lentogenic APMV-1 strains. Phylogenetic analysis of full-length F genes of all isolates was performed, and based on the recently proposed criteria for classification of APMV-1 strains, the 24 Colombian isolates were found to belong to class II viruses and clustered into four different genotypes. Ten virulent isolates clustered with genotype VII (sub-genotype VIId), seven lentogenic strains within genotype II, five lentogenic strains with genotype I (sub-genotype Ia), and two virulent isolates within genotype XII. Our data provide essential information on the genetic diversity of AMPV-1 isolates circulating in Colombia.

Novel pyrazolo[5,1-f][1,6]naphthyridines, pyrazolo[5,1-a][2,6]naphthyridines, pyrazolo[5,1-a][2,7]naphthyridines and pyrazolo[5,1-a]isoquinolines phenylimidazole/benzimidazole ethylene-linked were designed and synthesized for PDE10A interaction. An AgOTf and proline-cocatalyzed multicomponent methodology based on use of o-alkynylaldehydes, tosylhydrazide and ketones was developed and proved to be a convenient route for assembly of most of the novel tricyclic pyrazoles synthesized. Pyrazolo[5,1-f][1,6]naphthyridine 43 and 59, pyrazolo[5,1-a][2,6]naphthyridine 66, and pyrazolo[5,1-a][2,7]naphthyridine 42 showed the highest affinity for PDE10A enzyme (IC50 = 40, 42, 40, 55 nM, respectively).

The scientific advances during the 1970ies and 1980ies within the field of dopaminergic neurotransmission enabled the development of a pharmacophore that became the template for design and synthesis of dopamine D2 agonists during the following four decades. A major drawback, however, is that this model fails to accommodate certain classes of restrained dopamine D2 agonists including ergoline structures. To accommodate these, a revision of the original model was required. The present study has addressed this by an extension of the original model without compromising its obvious qualities. The revised pharmacophore contains an additional hydrogen bond donor feature, which is required for it to accommodate ergoline structures in a low energy conformation and in accordance with the steric restrictions dictated by the original model. The additional pharmacophore feature suggests ambiguity in the binding mode for certain compounds, including a series of ergoline analogues, which was reported recently. The ambiguity was confirmed by docking to a homology model of the D2 receptor as well as by pharmacological characterization of individual enantiomers of one of the analogues. The present research also addresses the potential of designing ligands that interact with the receptor in a large, distal cavity of the dopamine D2 receptor that has not previously been studied systematically. The pharmacological data indicate that this area may be a major determinant for both the dopamine D2 affinity and efficacy, which remains to be explored in future studies.

2,5-Dimethoxyphenethylamines and their N-benzylated derivatives are potent 5-HT2A agonists with psychedelic effects in humans. The N-benzylated derivatives are among the most selective 5-HT2A agonists currently available and their usage as biochemical and brain imaging tools is increasing, yet very little is known about the relationships between the structure of the ligands and their pharmacokinetic profile. In order to evaluate the potential of these compounds for in vivo applications we have determined the microsomal stability of 11 phenethylamines and 27 N-benzylated derivatives thereof using human liver microsomes. We found that the N-benzylated phenethylamines have much higher intrinsic clearance than the parent phenethylamines. We hypothesize that their low hepatic stability renders them orally inactive due to first pass metabolism, which is supported by anecdotal data from recreational use of these compounds.

While previous investigations have demonstrated the benefit of cardiac rehabilitation (CR) on outcomes after cardiac surgery, the association between pre-operative frailty and post-operative CR completion is unclear. The purpose of this retrospective cohort study was to determine if pre-operative frailty scores impacted CR completion post-operatively and if CR completion influenced frailty scores in 114 cardiac surgery patients. Frailty was assessed with the use of the Clinical Frailty Scale (CFS), the Modified Fried Criteria (MFC), the Short Physical Performance Battery (SPPB), and the Functional Frailty Index (FFI). A Mann-Whitney test was used to compare frailty scores between CR completers and non-completers and changes in frailty scores from baseline to 1-year post-operation. CR non-completers were more frail than CR completers at pre-operative baseline based on the CFS (p = 0.01), MFC (p &lt, 0.001), SPPB (p = 0.007), and the FFI (p &lt, 0.001). A change in frailty scores from baseline to 1-year post-operation was not detected in either group using any of the four frailty assessments. However, greater improvements from baseline to 1-year post-operation in two MFC domains (cognitive impairment and low physical activity) and the physical domain of the FFI were found in CR completers as compared to CR non-completers. These data suggest that pre-operative frailty assessments have the potential to identify participants who are less likely to attend and complete CR. The data also suggest that frailty assessment tools need further refinement, as physical domains of frailty function appear to be more sensitive to change following CR than other domains of frailty.

This study tested the hypothesis that exercise training would prevent the development of diabetes-induced cardiac dysfunction and altered expression of sarcoplasmic reticulum Ca2 +-transport proteins in the low-dose streptozotocin-induced diabetic rats fed a high-fat diet (HFD+STZ). Male Sprague–Dawley rats (4 weeks old; 125–150 g) were made diabetic using a high-fat diet (40% fat, w/w) and a low-dose of streptozotocin (35 mg·(kg body mass)–1) by intravenous injection. Diabetic animals were divided among a sedentary group (Sed+HFD+STZ) or an exercise-trained group (Ex+HFD+STZ) that accumulated 3554 ± 338 m·day–1of voluntary wheel running (mean ± SE). Sedentary animals fed a low-fat diet served as the control (Sed+LFD). Oral glucose tolerance was impaired in the sedentary diabetic group (1179 ± 29; area under the curve (a.u.c.)) compared with that in the sedentary control animals (1447 ± 42 a.u.c.). Although left ventricular systolic function was unchanged by diabetes, impaired E/A ratios (i.e., diastolic function) and rates of pressure decay (–dP/dt) indicated the presence of diastolic dysfunction. Diabetes also reduced SERCA2a protein content and maximal SERCA2a activity (Vmax) by 21% and 32%, respectively. In contrast, the change in each parameter was attenuated by exercise training. Based on these data, it appears that exercise training prevented the development of diabetic cardiomyopathy and the dysregulation of sarcoplasmic reticulum protein content in an inducible animal model of type 2 diabetes.

Inhibitors of the phosphodiesterase enzyme PDE10A have been the target for extensive investigations and huge drug discovery research efforts during the recent years. Although PDE10A with its 13 years history is a relatively newly discovered target, it has been paradigmatic for the new generation of 'high efficiency drug discovery'. Several companies now have clinical programs aiming at validating the clinical potential of PDE10A inhibitors. The majority of companies have been focusing on the treatment of schizophrenia since preclinical evidence suggests that a PDE10A inhibitor could provide antipsychotic, pro-cognitive and negative symptom efficacy.This article highlights and reviews research advances published in the patent literature since mid-2009 until mid-2012. The article is supplemented with selected publications from the scientific literature, emphasizing the possible involvement of PDE10A inhibitors in the treatment of schizophrenia.Several compounds from various companies are currently undergoing clinical testing, dominated by compounds in clinical Phase I. Focus is mainly on CNS diseases and schizophrenia is the leading target indication.

Purpose Muller cells are considered to be vital in the maintenance of retinal ganglion cells (RGCs), and since RGCs are essential to maintain the neuronal function of the retina, a functioning symbiotic partnership between Muller cells and RGCs is fundamental. The present study evaluates glia-neuron interactions in a coculture model of primary Muller cells and primary RGCs. Methods To investigate the Muller cell-RGC interaction we developed a coculture model, in which primary Muller cells from mice were grown in inserts on top of pure primary RGC cultures likewise from mice. The impact of 24 h of starvation on the ability of Muller cells to protect RGCs was evaluated. Moreover, changes in glutamate uptake were studied in Muller cells as well as cell viability in response to starvation. Results The presence of Muller cells significantly increased the survival of RGCs during normal conditions as well as during 24 h of pre-starvation. Glutamate uptake capacity was significantly increased during starvation along with an increased Vmax. Starvation induced significantly reduced cell viability both in Muller cells and RGCs with a significantly greater reduction of cell viability in Muller cells. Conclusions The present study reveals an increased survival of RGCs in presence of Muller cells and herewith we confirm Muller cells as being key players in RGC homeostasis. Moreover, we verify the coculture model as being a unique approach to study the glia-neuron partnership in inner retinal diseases.

Aim: Large-molecule biotherapeutic quantitation in vivo by LC–MS has traditionally relied on enzymatic digestion followed by quantitation of a ‘surrogate peptide’ to infer whole-molecule concentration. MS methods presented here measure the whole molecule and provide a platform to better understand the various circulating drug forms by allowing for variant quantitation. Results: An immunocapture LC–MS method for quantitation of a biotherapeutic monoclonal antibody from human plasma is presented. Sensitivity, precision and accuracy for each molecular portion are presented along with an example of glycoform variant quantitation. Conclusion: The method is presented as a basic platform to be further developed for Good Practice (GxP) applications, critical quality attribute analysis or general understanding of molecular forms present as required for the wide range of drug development processes.

Glia-neuron partnership is important for inner retinal homeostasis and any disturbances may result in retinal ganglion cell (RGC) death. Müller cells support RGCs with essential functions such as removing excess glutamate and providing energy sources. The aim was to explore the impact of Müller cells on RGC survival. To investigate the Müller cell/RGC interactions we developed a coculture model, in which primary Müller cells were grown in inserts on top of pure primary RGC cultures. The impact of starvation and mitochondrial inhibition on the Müller cell ability to protect RGCs was studied. Moreover, the ability of Müller cells to remove glutamate from the extracellular space was investigated. RGC survival was evaluated by cell viability assays and glutamate uptake was assessed by kinetic uptake assays. We demonstrated a significantly increased RGC survival in presence of untreated and prestarved Müller cells. Additionally, prestarved Müller cells significantly increased RGC survival after mitochondrial inhibition. Finally, we revealed a significantly increased ability to take up glutamate in starved Müller cells. Overall, our study confirms essential roles of Müller cells in RGC survival. We suggest that targeting Müller cell function could have potential for future treatment strategies to prevent blinding neurodegenerative retinal diseases.

We propose that pathological remodeling in joint tissues of osteoarthritis (OA) patients persistently stimulates local secretion of pro-inflammatory mediators, which overflow into the blood, activating leukocytes that impair endothelial function and accelerate the atherosclerotic process. During periods of pain, endothelial dysfunction progresses more aggressively due to elevated secretion of these pro-inflammatory mediators, which are involved in both atherosclerosis and the sensation of pain. Concentrations of pro-inflammatory cytokines and their antagonists, activating and decoy receptors of the broad interleukin (IL)-1 and IL-17 families, IL-15, and monocyte chemotactic protein-1 should be measured in peripheral blood samples of OA patients and compared with (I) OA clinical severity ; (II) subclinical parameters of atherosclerosis ; (III) ischemic heart disease risk factors ; (IV) soluble factors indicating endothelial dysfunction ; (V) degree of bone destruction ; and (VI) results of a six-minute walk test. Arthroscopy and joint replacement surgery provide an opportunity to estimate mRNA and protein expression of inflammatory mediators in specimens of synovial fluid, synovial membrane, cartilage, and/or subarticular bone. A range of methods, including questionnaires, X-ray, computed tomography, ultrasound, enzyme-linked immunosorbent assay, immunohistology, immunofluorescence, and reverse transcription and in situ polymerase chain reaction are available. Understanding the inflammatory and immune mechanisms underlying OA may allow the early identification of patients at high risk of cardiovascular disease, independently of classical coronary risk factors. Pain may constitute an extrinsic indicator of currently worsening endothelial function

Objective Shunt obstructions may partly be caused by brain debris, which intrude into the ventricular catheter during ventricle puncture. Avoiding contact between the catheter and brain tissue, by using a peel-away sheath, should reduce the number of shunt failures caused by obstruction. To test this hypothesis, we conducted a randomized, prospective multicenter study. Methods 201 patients from 6 different neurosurgical centers in Germany receiving a ventriculo-peritoneal shunt were included in this study. Of these, 177 patients completed a 1-year follow-up period. Surgery was randomized in a 1 to 1 fashion, such that out of 177 procedures, 91 were performed using a peel-away sheath and 86 were performed without. The rate of surgical re-interventions and shunt obstructions within a 12-month period was recorded. Results Within 1 year post-surgery, 17 shunt obstructions (9.6%) leading to shunt revisions were recorded. However, no difference was found between surgeries performed using a peel-away sheath (9.9%) or not (9.3%). The overall shunt infection rate was 2.8% and the shunt revision rate for overdrainage was 3.9%. Conclusions The theoretical advantages attributed to the use of a peel-away sheath to introduce a ventricular catheter could not be confirmed in this randomized study, suggesting that the proposed role of brain debris in shunt obstructions may be overestimated.

Background: The quantification of proteins and peptides in in vivo samples is a critical part of supporting the drug development process for biotherapeutics. LC–MS/MS using tandem quadrupole mass spectrometers is well established as the technology of choice for the quantification of small-molecule drugs and their metabolites in biological fluid. The application of accurate mass MS for quantification in a DMPK environment has attracted considerable interest in recent years. Materials & Methods: In this article we describe and compare the application of LC–high-resolution MS and LC–selected reaction monitoring (SRM) for the quantification of a therapeutics proteins. Results: The accurate mass instrumentation showed acceptable linearity and sensitivity to quantify the protein therapeutic to the level of 10 ng/ml. The accurate mass instrument was operated in accurate mass SRM using high resolution (SRM-HR), the assay was demonstrated to be linear over three orders of magnitude. By narrowing the mass window from 100 mDa to 40 mDa and then to 20 mDa the assay specificity was significantly improved, hence increasing the S/N and improving the assay sensitivity. Conclusion: The high-resolution instrument was demonstrated to be reproducible over the course of the assay. The accurate mass method sensitivity was determined to be within one order of magnitude of that obtained with a tandem quadrupole MS/MS assay.

Object Ventriculoatrial (VA) shunts inserted for the treatment of hydrocephalus are known to be a risk factor for pulmonary hypertension. The aim of this study was to evaluate the incidence of pulmonary hypertension among adult patients with VA shunts. Methods All patients who had received a VA shunt at one of two institutions between 1985 and 2000 were invited for a cardiopulmonary evaluation. The investigation included a thorough history taking, clinical examination, echocardiography, and pulmonary function testing including diffusing capacity of the lung for carbon monoxide (DLCO). Pulmonary hypertension was defined as systolic pulmonary artery pressure > 35 mm Hg at rest. Results The study group consisted of 86 patients, of whom 38 (44%) could be examined. The patients' mean age was 47.1 ± 18.4 years; the median interval between shunt insertion and cardiopulmonary evaluation was 15 years (range 5–20 years). Of the 38 patients, 20 (53%) had Doppler velocity profiles of tricuspid regurgitation that were adequate for the estimation of pulmonary artery systolic pressure. Doppler-defined pulmonary hypertension was observed in 3 patients (8%), 2 of whom underwent right heart catheterization. Chronic thromboembolic pulmonary hypertension was confirmed in both patients, and medical therapy, including anticoagulation, was started. The VA shunt was removed in both cases and replaced with a different type of device. Pulmonary function tests revealed a restrictive pattern in 15% and typical obstructive findings in 9% of patients. In 30% of patients the DLCO was less than 80% of predicted, and blood gas analysis showed hypoxemia in 6% of patients. No significant differences in pulmonary function tests were noted between the patients with and without echocardiographic evidence of pulmonary hypertension. However, patients with pulmonary hypertension had significantly lower DLCO values. Conclusions The authors detected pulmonary hypertension by using Doppler echocardiography in a significant proportion of patients with VA shunts. It is therefore recommended that practitioners perform regular echocardiography and pulmonary function tests, including single-breath DLCO in these patients to screen for pulmonary hypertension to prevent hazardous late cardiopulmonary complications.