Recommendations for high intensity upper body exercise testing

Introduction: For given submaximal and maximal peak power outputs aerobic responses to upper body exercise are different to those for lower body exercise (Sawka, 1986: Exercise & Sport Sciences Reviews, 14, 175-211). However, much less is known regarding responses to exercise intensities at and around peak oxygen up take (VO2peak). Purpose: The purpose of this study was to determine the metabolic responses during arm crank ergometry (ACE) below, at and above peak oxygen uptake and to help establish exercise testing guidelines for high intensity upper body exercise. Methods: Following institutional ethical approval fourteen male students (Age 21.1, s = 6.1 years and 2.44 s=0.44 VO2peak) volunteered to take part in this study. Each participant exercised on a table mounted cycle ergometer (Monark 894E, Monark Exercise AB, Sweden). After habituation peak minute power (PMP) was calculated from an incremental test. Subsequently each participant completed four continuous work tests (CWT) to volitional exhaustion at 80%, 90%, 100% and 110% of PMP. All tests were completed at 70 rev∙min-1 with a minimum of 48-h between tests and the order was counterbalanced. Each CWT was preceded by a 5 min warm-up, loaded with a mass corresponding to the participants 80% PMP for 20 s at minutes 2, 3 and 4. Oxygen uptake (VO2), respiratory exchange ratio (RER), heart rate (HR) and ratings of perceived exertion for the arms (local (RPEL) and cardiorespiratory strain (RPECR) were recorded at 1 min, 2 min and at volitional exhaustion. The EMG responses at three sites (flexor carpi ulnaris, biceps brachii and triceps brachii lateral) were recorded using double-differential (16-3000 Hz bandwidth, x300 gain), bipolar, active electrodes (MP-2A, Linton, Norfolk, UK). Electromyographic data were sampled at 1000 Hz and filtered using a 20 to 500 Hz band-pass filter (MP150 Data Acquisition and AcqKnowledge 4.0, Biopac, Goleta, CA). The EMG signals for each muscle were root mean squared (RMS) with a 500-ms sample window. The signal was then normalised, prior to each CWT, as a percentage of the mean of 3 sets of 10 duty cycles completed during the warm-up (see above) when the participants 80% PMP for 20 s was applied. Time to exhaustion (Tlim) was recorded as the performance outcome measure. Data for Tlim were analysed using one-way analysis of variance. Differences in EMG, VO2, RER, HR, RPEL and RPECR were analysed using separate two-way analysis of variance with repeated measures (trial x time). All analyses were performed using the Statistical Package for Social Sciences ( 17.0; SPSS Inc., Chicago, IL). Individual differences in means were located using Bonferroni post-hoc correction. Significance was accepted at P < 0.05. Results: As resistive load increased Tlim decreased (611 s=194, 397 s=99, 268 s=90, 206 s=67s, respectively; P < 0.001, ES = 0.625). Post-hoc analysis revealed that Tlim using 80%PMP was longer than for 90%, 100% and 110% PMP trials (P < 0.001) and 90% was longer than both 100% and 110% PMP trials (P = 0.079, P = 0.001). At exhaustion VO2 was similar across trials (P = 0.413, ES = 0.053), although 80% PMP VO2 tended to be less (2.10 s=0.32 l·min-1) than for 90% (2.29 s=0.37), 100% (2.33 s=0.49) and 110% (2.26 s=0.34). Also, 80% PMP VO2 was less than VO2peak (P = 0.013). There were differences in RER at Tlim (P < 0.001, ES = 0.593) with values increasing with % PMP (1.15 s=0.07, 1.26 s=0.07, 1.36 s=0.10, 1.40 s=0.09, respectively). There were no differences across trials for HR at Tlim (~173 (12); P = 0.834, ES = 0.016) and HR was proportional to %PMP at 1 min, and 2 min. For flexor carpi ulnaris there was an increase in activation as exercise intensity increased (P < 0.001, ES = 0.245). There were a similar responses for biceps brachii and triceps brachii demonstrating an increase in activation with exercise intensity (P <0.001, ES = 0.137, P < 0.001, ES = 0.163, respectively). No differences for RPEL and RPECR were observed at Tlim. Discussion: There was a clear response of Tlim with intensity as expected for lower body exercise (Hill et al., 2002: Medicine and Science in Sports and Exercise, 34(4), 709-714). Despite differences in Tlim across exercise intensities VO2, HR and RPE were similar at exhaustion indicating a functional cardiorespiratory maximum had been reached. As indicated by the RER an increased activation of the anaerobic metabolism with greater exercise intensities (100% and 110%) is likely and therefore this may represent a greater anaerobic component at these two intensities. The increase in EMG activity with intensity could indicate an increase activity with an increase in exercise intensity. Conclusion: It is recommended that due to the combination of muscle activation, oxygen uptake and Tlim that an exercise intensity of 90% or 100% of PMP could be used for high intensity upper body exercise testing

Application of ER Stress Biomarkers to Predict Formulated Monoclonal Antibody Stability

For a therapeutic mAb to reach the clinic, the molecule must be produced at an appropriate yield and quality, then formulated to maintain efficacy and stability. The formation of sub‐visible particles (SVPs) can impact on product stability and is monitored during formulation development however, the potential of a mAb to form such species can be influenced throughout the whole bioprocess. We investigate levels of intracellular ER stress perceived by cells, day of mAb harvest and the relationship to subsequent product stability of two mAbs (denoted A and B), produced in CHO cell lines, as determined by SVP content after accelerated stability studies. We show the propensity of mAb A to form SVPs can be predicted by transcript expression of biomarkers of cellular ER stress, heavy/light chain transcript and polypeptide amounts, and harvest day. Further, mAb A material harvested on day 9 of culture was more stable, in terms of SVP formation, than material harvested on day 13. These data suggest that ER stress perceived by CHO cells during culture can reflect the stability of a mAb, and that biomarkers of such stress could help define culture harvest time as a tool to control or reduce SVP formation in formulated mAbs

An Influenza Virus M2 Protein Specific Chimeric Antigen Receptor Modulates Influenza A/WSN/33 H1N1 Infection In Vivo

A potential target for the development of universal vaccine strategies against Influenza A is the M2 protein – a membrane protein with a highly conserved extracellular domain. In this study we developed engineered T-cell receptors, by fusing M2-specific antibody sequences with T-cell receptor transmembrane and signaling domains to target influenza infected cells. When expressed on T-cells, these novel T-cell receptors (chimeric antigen receptors - CARs) are able to recognize specific antigens on the surface of target cells via an MHC-independent mechanism. Using an existing monoclonal antibody (14C2) specific for the M2 ectodomain (M2e), we generated an M2-specific CAR. We tested the specificity of this M2 CAR in vitro by measuring the activation of T-cells in response to M2-specific peptides or M2-expressing cell lines. Both Jurkat T-cells and peripheral blood mononuclear cells expressing the M2-specific CAR responded to specific antigen stimulation by upregulating NFAT and producing γ-interferon. To test whether the M2-specific CAR are effective at recognizing influenza infected cells in vivo we used an established BALB/c murine infection model. At day 4 post-infection, when M2 CAR expressing splenocytes could be detected in the lung, the Influenza A/WSN/33 virus titre was around 50% of that in control mice. Although the lung virus titre later increased in the treated group, virus was cleared in both groups of mice by day 8. The results provide support for the development of M2e as a target for cell mediated immunotherapy

MMC Fall with Injury Prevention Project

Problem/Impact Statement: Patients falls with injury remains an elusive problem at MMC. Over the past 8 quarter, (2016 and 2017) MMC has outperformed 3 of the last 8 Quarters of data. The average rate for the past 8 quarters is .57/1000 patient days with the mean benchmark of .54/per 1000 patient days. MH has determined a focus goal for all the MH hospitals to be below .70/MH 100 patient days as a goal for falls with injury. MMC having the largest volume must be below NDNQI mean to drive this change as the .70 is the average of all MH hospitals. A fall with injury costs on Average cost of a fall with injury is $14,000., more importantly the cost to the patient may be an increase in hospital stay, and increase in level of care. Injuries range from lacerations to fractures and head trauma and death. Approximately 50% of all falls incur an injury. Putting interventions in place to decrease total falls will decrease injuries at MMC

Designing a CRISPR-Cas9 pipeline to investigate the effects of putative genetic modifiers on Duchenne Muscular Dystrophy (DMD)

Duchenne muscular dystrophy (DMD) is an X-linked neuromuscular disorder that affects about 1 in 3,500 live male births (Bushby et al. 2010). DMD is caused by a mutation of the Dystrophin (or DMD) gene that causes loss of dystrophin, a structural protein. Loss of dystrophin increases muscle's susceptibility to damage leading to muscle atrophy and loss of muscle function (Bass et al. 2016). Disease progression is variable in patients with DMD. Genome-wide association studies (GWAS) have identified putative genetic modifiers of DMD that may influence disease severity and variability (Bello et al. 2016; Flangian et al. 2013; Flanigan et al. 2021; Wess et al. 2018). I am using zebrafish, an established model organism to study DMD, to validate and further investigate these putative modifiers. CRISPR-Cas9 mutagenesis is highly efficient in zebrafish and can be used for rapid genetic screens by injecting Cas9 protein and a gene-specific sgRNA, targeting vital protein domains, to knock-out gene function and generate crispant fish. I designed sgRNAs to target zebrafish orthologs of DMD, LTBP4, THBS1, ETAA1, and PARD6G. Microinjection of zebrafish embryos with Cas9 and sgRNA targeting dmd causes loss of dystrophin protein, disorganization of muscle when assayed by birefringence, impaired motility, and decreased life-span of injected fish. To assay the effect of these genetic modifiers on dmd, multiplexed injections of dmd sgRNA + sgRNA targeting putative modifiers were carried out. To validate mutagenesis of putative modifiers hRMA was conducted after injection. Fish that show a dmd phenotype and mutagenesis of the putative modifier can then be used for subsequent analysis of muscle birefringence, motility, and lifespan. My data lays the foundational work to rapidly screen and identify modifiers of DMD for the development of new DMD therapeutics.Undergraduate Research Apprenticeship ProgramNationwide Children's HospitalOSU President's Research Excellence Accelerator AwardAcademic Major: Biolog

A collection of open problems in celebration of Imre Leader's 60th birthday

One of the great pleasures of working with Imre Leader is to experience his infectious delight on encountering a compelling combinatorial problem. This collection of open problems in combinatorics has been put together by a subset of his former PhD students and students-of-students for the occasion of his 60th birthday. All of the contributors have been influenced (directly or indirectly) by Imre: his personality, enthusiasm and his approach to mathematics. The problems included cover many of the areas of combinatorial mathematics that Imre is most associated with: including extremal problems on graphs, set systems and permutations, and Ramsey theory. This is a personal selection of problems which we find intriguing and deserving of being better known. It is not intended to be systematic, or to consist of the most significant or difficult questions in any area. Rather, our main aim is to celebrate Imre and his mathematics and to hope that these problems will make him smile. We also hope this collection will be a useful resource for researchers in combinatorics and will stimulate some enjoyable collaborations and beautiful mathematics

Improving Safe Handoffs & Transitions from the ED to Adult Inpatient: A Response to the AHRQ Hospital Patient Safety Culture Survey

SAFE TRANSITIONS AND PATIENT HANDOFFS IN A LARGE ACUTE CARE HOSPITAL It is well documented in the literature that ineffective patient handoffs and transitions continues to be an area that can lead to adverse patient safety events so it is an urgent opportunity for a performance improvement plan. At an academic tertiary care medical center, the lowest scoring domain from the FY2017 AHRQ Patient Safety Culture Survey was patient handoffs and transitions. A team was established consisting of staff from the Emergency Department and a medical/surgical unit to develop a plan for implementing improvement interventions. Their goal was to attain a ≥ 5% improvement on the patient handoff and transitions question in the Survey by the end of 12/30/2018. A root cause analysis was initiated and resulted in a KPI being developed to collect baseline data on handoffs and transitions between the two units. After three phases to analyze and improve, several countermeasures were created. Amongst them was the development of a evidence based mnemonic handoff tool to be used within 30 minutes of patient transfer. Pulse surveys of the involved units conducted at various intervals demonstrated significant improvement of \u3e5% in the 3 teamwork questions. Next steps will be to standardize, sustain and spread the improvement methods

Stratification of PD-1 blockade response in melanoma using pre- and post-treatment immunophenotyping of peripheral blood

Efficacy of checkpoint inhibitor therapies in cancer varies greatly, with some patients showing complete responses while others do not respond and experience progressive disease. We aimed to identify correlates of response and progression following PD-1-directed therapy by immunophenotyping peripheral blood samples from 20 patients with advanced malignant melanoma before and after treatment with the PD-1 blocking antibody pembrolizumab. Our data reveal that individuals responding to PD-1 blockade were characterised by increased CD8 T cell proliferation following treatment, while progression was associated with an increase in CTLA-4-expressing Treg. Remarkably, unsupervised clustering analysis of pre-treatment T cell subsets revealed differences in individuals that went on to respond to PD-1 blockade compared to individuals that did not. These differences mapped to expression of the proliferation marker Ki67 and the costimulatory receptor CD28 as well as the inhibitory molecules 2B4 and KLRG1. While these results require validation in larger patient cohorts, they suggest that flow cytometric analysis of a relatively small number of T cell markers in peripheral blood could potentially allow stratification of PD-1 blockade treatment response prior to therapy initiation

Development and validation of a high density SNP genotyping array for Atlantic salmon (Salmo salar)

BackgroundDense single nucleotide polymorphism (SNP) genotyping arrays provide extensive information on polymorphic variation across the genome of species of interest. Such information can be used in studies of the genetic architecture of quantitative traits and to improve the accuracy of selection in breeding programs. In Atlantic salmon (Salmo salar), these goals are currently hampered by the lack of a high-density SNP genotyping platform. Therefore, the aim of the study was to develop and test a dense Atlantic salmon SNP array. ResultsSNP discovery was performed using extensive deep sequencing of Reduced Representation (RR-Seq), Restriction site-Associated DNA (RAD-Seq) and mRNA (RNA-Seq) libraries derived from farmed and wild Atlantic salmon samples (n = 283) resulting in the discovery of &gt; 400 K putative SNPs. An Affymetrix Axiom&reg; myDesign Custom Array was created and tested on samples of animals of wild and farmed origin (n = 96) revealing a total of 132,033 polymorphic SNPs with high call rate, good cluster separation on the array and stable Mendelian inheritance in our sample. At least 38% of these SNPs are from transcribed genomic regions and therefore more likely to include functional variants. Linkage analysis utilising the lack of male recombination in salmonids allowed the mapping of 40,214 SNPs distributed across all 29 pairs of chromosomes, highlighting the extensive genome-wide coverage of the SNPs. An identity-by-state clustering analysis revealed that the array can clearly distinguish between fish of different origins, within and between farmed and wild populations. Finally, Y-chromosome-specific probes included on the array provide an accurate molecular genetic test for sex. ConclusionsThis manuscript describes the first high-density SNP genotyping array for Atlantic salmon. This array will be publicly available and is likely to be used as a platform for high-resolution genetics research into traits of evolutionary and economic importance in salmonids and in aquaculture breeding programs via genomic selection