A new strategy for faster urinary biomarkers identification by Nano-LC-MALDI-TOF/TOF mass spectrometry

<p>Abstract</p> <p>Background</p> <p>LC-MALDI-TOF/TOF analysis is a potent tool in biomarkers discovery characterized by its high sensitivity and high throughput capacity. However, methods based on MALDI-TOF/TOF for biomarkers discovery still need optimization, in particular to reduce analysis time and to evaluate their reproducibility for peak intensities measurement. The aims of this methodological study were: (i) to optimize and critically evaluate each step of urine biomarker discovery method based on Nano-LC coupled off-line to MALDI-TOF/TOF, taking full advantage of the dual decoupling between Nano-LC, MS and MS/MS to reduce the overall analysis time; (ii) to evaluate the quantitative performance and reproducibility of nano-LC-MALDI analysis in biomarker discovery; and (iii) to evaluate the robustness of biomarkers selection.</p> <p>Results</p> <p>A pool of urine sample spiked at increasing concentrations with a mixture of standard peptides was used as a specimen for biological samples with or without biomarkers. Extraction and nano-LC-MS variabilities were estimated by analyzing in triplicates and hexaplicates, respectively. The stability of chromatographic fractions immobilised with MALDI matrix on MALDI plates was evaluated by successive MS acquisitions after different storage times at different temperatures.</p> <p>Low coefficient of variation (CV%: 10–22%) and high correlation (R²> 0.96) values were obtained for the quantification of the spiked peptides, allowing quantification of these peptides in the low fentomole range, correct group discrimination and selection of "specific" markers using principal component analysis. Excellent peptide integrity and stable signal intensity were found when MALDI plates were stored for periods of up to 2 months at +4°C. This allowed storage of MALDI plates between LC separation and MS acquisition (first decoupling), and between MS and MSMS acquisitions while the selection of inter-group discriminative ions is done (second decoupling). Finally the recording of MSMS spectra to obtain structural information was focused only on discriminative ions in order to minimize analysis time.</p> <p>Conclusion</p> <p>Contrary to other classical approaches with direct online coupling of chromatographic separation and on the flight MS and/or MSMS data acquisition for all detected analytes, our dual decoupling strategy allowed us to focus on the most discriminative analytes, giving us more time to acquire more replicates of the same urine samples thus increasing detection sensitivity and mass precision.</p

Evidence of disturbed sleep and increased illness in overreached endurance athletes

PURPOSE: This study aimed to examine whether (i) objective markers of sleep quantity and quality are altered in endurance athletes experiencing overreaching in response to an overload training program and (ii) potential reduced sleep quality would be accompanied with a higher prevalence of upper respiratory tract infections in this population. METHODS: Twenty-seven trained male triathletes were randomly assigned to either overload (n = 18) or normal (CTL, n = 9) training groups. Respective training programs included a 1-wk moderate training phase followed by a 3-wk period of overload or normal training, respectively, and then a subsequent 2-wk taper. Maximal aerobic power and oxygen uptake (V̇O 2max) from incremental cycle ergometry were measured after each phase, whereas mood states and incidences of illness were determined from questionnaires. Sleep was monitored every night of the 6 wk using wristwatch actigraphy. RESULTS: Of the 18 overload training group subjects, 9 were diagnosed as functionally overreached (F-OR) after the overload period, as based on declines in performance and V̇O2max with concomitant high perceived fatigue (P 0.05). There was a significant time-group interaction for sleep duration (SD), sleep efficiency (SE), and immobile time (IT). Only the F-OR group demonstrated a decrease in these three parameters (-7.9% ± 6.7%,-1.6% ± 0.7%, and-7.6% ± 6.6% for SD, SE, and IT, respectively, P < 0.05), which was reversed during the subsequent taper phase. Higher prevalence of upper respiratory tract infections were also reported in F-OR (67%, 22%, and 11% incidence rate for F-OR, AF, and CTL, respectively). CONCLUSION: This study confirms sleep disturbances and increased illness in endurance athletes who present with symptoms of F-OR during periods of high volume training. © 2014 by the American college of Sports Medicine

Heat-acclimatization and pre-cooling: a further boost for endurance performance?

© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd To determine if pre-cooling (PC) following heat-acclimatization (HA) can further improve self-paced endurance performance in the heat, 13 male triathletes performed two 20-km cycling time-trials (TT) at 35 °C, 50% relative humidity, before and after an 8-day training camp, each time with (PC) or without (control) ice vest PC. Pacing strategies, physiological and perceptual responses were assessed during each TT. PC and HA induced moderate (+10 ± 18 W; effect size [ES] 4.4 ± 4.6%) and very large (+28 ± 19 W; ES 11.7 ± 4.1%) increases in power output (PO), respectively. The overall PC effect became unclear after HA (+4 ± 14 W; ES 1.4 ± 3.0%). However, pacing analysis revealed that PC remained transiently beneficial post-HA, i.e., during the first half of the TT. Both HA and PC pre-HA were characterized by an enhanced PO without increased cardio-thermoregulatory or perceptual disturbances, while post-HA PC only improved thermal comfort. PC improved 20-km TT performance in unacclimatized athletes, but an 8-day HA period attenuated the magnitude of this effect. The respective converging physiological responses to HA and PC may explain the blunting of PC effectiveness. However, perceptual benefits from PC can still account for the small alterations to pacing noted post-HA

The 2mrad crossing angle scheme for the international linear collider

http://cern.ch/AccelConf/e08/papers/mopp005.pdfInternational audienceThe present baseline configuration of the ILC has a 14 mrad crossing angle between the beams at the interaction point. This allows easier extraction of the beams after col- lisions, but imposes on the other hand more constraints on the control of the beams prior to colliding them. More- over, some limitations to physics capabilities arise, in par- ticular because of the degraded very forward electromag- netic detector hermeticity and because calibration proce- dures for (gaseous) tracking detectors become more com- plex. To mitigate these problems, alternative configurations with very small crossing angles are studied. A new version of the 2 mrad layout was designed last year, based on sim- pler concepts and assumptions. The emphasis of this new scheme was to satisfy specifications with as few and feasi- ble magnets as possible, in order to reduce costs

The Development of Functional Overreaching Is Associated with a Faster Heart Rate Recovery in Endurance Athletes

Purpose The aim of the study was to investigate whether heart rate recovery (HRR) may represent an effective marker of functional overreaching (f-OR) in endurance athletes. Methods and Results Thirty-one experienced male triathletes were tested (10 control and 21 overload subjects) before (Pre), and immediately after an overload training period (Mid) and after a 2-week taper (Post). Physiological responses were assessed during an incremental cycling protocol to exhaustion, including heart rate, catecholamine release and blood lactate concentration. Ten participants from the overload group developed signs of f-OR at Mid (i.e. -2.1 ± 0.8% change in performance associated with concomitant high perceived fatigue). Additionally, only the f-OR group demonstrated a 99% chance of increase in HRR during the overload period (+8 ± 5 bpm, large effect size). Concomitantly, this group also revealed a >80% chance of decreasing blood lactate (-11 ± 14%, large), plasma norepinephrine (-12 ± 37%, small) and plasma epinephrine peak concentrations (-51 ± 22%, moderate). These blood measures returned to baseline levels at Post. HRR change was negatively correlated to changes in performance, peak HR and peak blood metabolites concentrations. Conclusion These findings suggest that i) a faster HRR is not systematically associated with improved physical performance, ii) changes in HRR should be interpreted in the context of the specific training phase, the athletes perceived level of fatigue and the performance response; and, iii) the faster HRR associated with f-OR may be induced by a decreased central commandand by a lower chemoreflex activity

Unified Image and Video Saliency Modeling

Visual saliency modeling for images and videos is treated as two independent tasks in recent computer vision literature. While image saliency modeling is a well-studied problem and progress on benchmarks like SALICON and MIT300 is slowing, video saliency models have shown rapid gains on the recent DHF1K benchmark. Here, we take a step back and ask: Can image and video saliency modeling be approached via a unified model, with mutual benefit? We identify different sources of domain shift between image and video saliency data and between different video saliency datasets as a key challenge for effective joint modelling. To address this we propose four novel domain adaptation techniques - Domain-Adaptive Priors, Domain-Adaptive Fusion, Domain-Adaptive Smoothing and Bypass-RNN - in addition to an improved formulation of learned Gaussian priors. We integrate these techniques into a simple and lightweight encoder-RNN-decoder-style network, UNISAL, and train it jointly with image and video saliency data. We evaluate our method on the video saliency datasets DHF1K, Hollywood-2 and UCF-Sports, and the image saliency datasets SALICON and MIT300. With one set of parameters, UNISAL achieves state-of-the-art performance on all video saliency datasets and is on par with the state-of-the-art for image saliency datasets, despite faster runtime and a 5 to 20-fold smaller model size compared to all competing deep methods. We provide retrospective analyses and ablation studies which confirm the importance of the domain shift modeling. The code is available at https://github.com/rdroste/unisalComment: Presented at the European Conference on Computer Vision (ECCV) 2020. R. Droste and J. Jiao contributed equally to this work. v3: Updated Fig. 5a) and added new MTI300 benchmark results to supp. materia

On blocks of defect two and one simple module, and Lie algebra structure of HH¹

Let k be a field of odd prime characteristic p. We calculate the Lie algebra structure of the first Hochschild cohomology of a class of quantum complete intersections over k. As a consequence, we prove that if B is a defect 2-block of a finite group algebra $kG$ whose Brauer correspondent C has a unique isomorphism class of simple modules, then a basic algebra of B is a local algebra which can be generated by at most 2√I elements, where I is the inertial index of B, and where we assume that k is a splitting field for B and C

Tacrolimus Population Pharmacokinetic-Pharmacogenetic Analysis and Bayesian Estimation in Renal Transplant Recipients

Objectives: The aims of this study were (i) to investigate the population pharmacokinetics of tacrolimus in renal transplant recipients, including the influence of biological and pharmacogenetic covariates; and (ii) to develop a Bayesian estimator able to reliably estimate the individual pharmacokinetic parameters and inter-dose area under the blood concentration-time curve (AUC) from 0 to 12 hours (AUC12) in renal transplant patients. Methods: Full pharmacokinetic profiles were obtained from 32 renal transplant patients at weeks 1 and 2, and at months 1, 3 and 6 post-transplantation. The population pharmacokinetic analysis was performed using the nonlinear mixed-effect modelling software NONMEM® version VI. Patients’ genotypes were characterized by allelic discrimination for PXR −25385C&gt;T genes. Results: Tacrolimus pharmacokinetics were well described by a two-compartment model combined with an Erlang distribution to describe the absorption phase, with low additive and proportional residual errors of 1.6 ng/mL and 9%, respectively. Both the haematocrit and PXR −25385C&gt;T single nucleotide polymorphism (SNP) were identified as significant covariates for apparent oral clearance (CL/F) of tacrolimus, which allowed improvement of prediction accuracy. Specifically, CL/F decreased gradually with the number of mutated alleles for the PXR −25385C&gt;T SNP and was inversely proportional to the haematocrit value. However, clinical criteria of relevance, mainly the decrease in interindividual variability and residual error, led us to retain only the haematocrit in the final model. Maximum a posteriori Bayesian forecasting allowed accurate prediction of the tacrolimus AUC12 using only three sampling times (at 0 hour [predose] and at 1 and 3 hours postdose) in addition to the haematocrit value, with a nonsignificant mean AUC bias of 2% and good precision (relative mean square error = 11%). Conclusion: Population pharmacokinetic analysis of tacrolimus in renal transplant recipients showed a significant influence of the haematocrit on its CL/F and led to the development of a Bayesian estimator compatible with clinical practice and able to accurately predict tacrolimus individual pharmacokinetic parameters and the AUC12

Clinical spectrum, prognosis and estimated prevalence of DNAJB11-kidney disease

Monoallelic mutations of DNAJB11 were recently described in seven pedigrees with atypical clinical presentations of autosomal dominant polycystic kidney disease. DNAJB11 encodes one of the main cofactors of the endoplasmic reticulum chaperon BiP, a heat-shock protein required for efficient protein folding and trafficking. Here we conducted an international collaborative study to better characterize the DNAJB11-associated phenotype. Thirteen different loss-of-function variants were identified in 20 new pedigrees (54 affected individuals) by targeted next-generation sequencing, whole-exome sequencing or whole-genome sequencing. Amongst the 77 patients (27 pedigrees) now in total reported, 32 reached end stage kidney disease (range, 55-89 years, median age 75); without a significant difference between males and females. While a majority of patients presented with non-enlarged polycystic kidneys, renal cysts were inconsistently identified in patients under age 45. Vascular phenotypes, including intracranial aneurysms, dilatation of the thoracic aorta and dissection of a carotid artery were present in four pedigrees. We accessed Genomics England 100,000 genomes project data, and identified pathogenic variants of DNAJB11 in nine of 3934 probands with various kidney and urinary tract disorders. The clinical diagnosis was cystic kidney disease for eight probands and nephrocalcinosis for one proband. No additional pathogenic variants likely explaining the kidney disease were identified. Using the publicly available GnomAD database, DNAJB11 genetic prevalence was calculated at 0.85/10.000 individuals. Thus, establishing a precise diagnosis in atypical cystic or interstitial kidney disease is crucial, with important implications in terms of follow-up, genetic counseling, prognostic evaluation, therapeutic management, and for selection of living kidney donors