On-the-fly rapid immunoassay for neonatal sepsis diagnosis: C-reactive protein accurate determination using magnetic graphene-based micromotors

Based on the exceptional and new opened biosensing possibilities of self-propelled micromotors, a micromotorbased immunoassay (MIm) has smartly been designed for C-reactive protein (CRP) determination in plasma of preterm infants with sepsis suspicion. The design of the micromotors involved the electrosynthesis of a carbon-based outer layer (for antibody functionalization), an intermediate Ni layer (for magnetic guidance and stopped flow operations) and PtNPs inner catalytic layer (for catalytic bubble propulsion). Micromotors biofunctionalization on the outer layer (using carbon black (CB), reduced graphene oxide (rGO) and multi-walled carbon nanotubes (MWCNTs), and biocompatible propulsion capabilities, were carefully studied. Magnetic rGO/Ni/PtNPs micromotors exhibited the most efficient and reproducible (CV ? 9%) anti-CRP functionalization, controlled stopped-flow operations as well as efficient bubble propulsion (1% H2O2, 1,5% NaCh, speed 140 ?m s-1). Analytical performance of MIm was excellent, allowing the direct (without dilution), sensitive (LOD ? 0.80 ?g/mL), and accurate CRP determination (Er ? 1%) in hardly available preterm babies? plasma samples with suspected sepsis using very low volumes (<10 ?L) and in just 5 min of on-the-fly bioassay. Overall, the results obtained allowed the fast and reliable sepsis diagnostics in preterm babies? individuals with suspected sepsis, not only proving the usefulness of the approach as its potential utilization as point-of-care device for clinical analysis but drawing new horizons in extremely low sample volumes-based diagnostics

New insights into the genetic component of non-infectious uveitis through an Immunochip strategy

Background: Large-scale genetic studies have reported several loci associated with specific disorders involving uveitis. Our aim was to identify genetic risk factors that might predispose to uveitis per se, independent of the clinical diagnosis, by performing a dense genotyping of immune-related loci. Methods: 613 cases and 3693 unaffected controls from three European case/control sets were genotyped using the Immunochip array. Only patients with non-infectious non-anterior uveitis and without systemic features were selected. To perform a more comprehensive analysis of the human leucocyte antigen (HLA) region, SNPs, classical alleles and polymorphic amino acid variants were obtained via imputation. A meta-analysis combining the three case/control sets was conducted by the inverse variance method. Results: The highest peak belonged to the HLA region. A more detailed analysis of this signal evidenced a strong association between the classical allele HLA-A*2902 and birdshot chorioretinopathy (p=3.21E-35, OR=50.95). An omnibus test yielded HLA-A 62 and 63 as relevant amino acid positions for this disease. In patients with intermediate and posterior uveitis, the strongest associations belonged to the rs7197 polymorphism, within HLA-DRA (p=2.07E-11, OR=1.99), and the HLA-DR15 haplotype (DRB1*1501: p=1.16E-10, OR=2.08; DQA1*0102: p=4.37E-09, OR=1.77; DQB1*0602: p=7.26E-10, OR=2.02). Outside the HLA region, the MAP4K4/IL1R2 locus reached statistical significance (rs7608679: p=8.38E-07, OR=1.42). Suggestive associations were found at five other loci. Conclusions: We have further interrogated the association between the HLA region and non-infectious non-anterior uveitis. In addition, we have identified a new non-HLA susceptibility factor and proposed additional risk loci with putative roles in this complex condition

Polymer-based micromotors fluorescence immunoassay for on the move sensitive procalcitonin determination in very low birth weight infants' plasma

A new fluorescence micromotor-based immunoassay (FMIm) has been developed forvprocalcitonin (PCT) determination as an early sepsis diagnostic analytical tool. The micromotorsvcombine the high binding capacity of the specific antibodies onto their polymeric polypyrrole outervlayer (PPy layer), with their magnetic guidance (Ni layer) and self-propulsion by catalytic generationvof oxygen bubbles (PtNP inner layer) to actively recognize the PCT antigen. This FMIm allowed avsensitive (LOD = 0.07 ng mL?1) and direct PCT determination in clinical samples from very lowbirth-vweight infants (VLBWI) with sepsis suspicion, using small volumes of sample (25 ?L) in a clinically relevant range of concentrations (0.5?150 ng mL?1). The good agreement between PCTvlevels obtained by our micromotor-based method and routine immunofluorescence hospitalvdetermination demonstrates the feasibility for the analysis in VLBWI samples and its potential as avpoint-of-care diagnostic tool for sepsis.Ministerio de Economía, Industria y Competitividad); Comunidad de Madrid; La Caix

Toward early diagnosis of late-onset sepsis in preterm neonates: Dual magnetoimmunosensor for simultaneous procalcitonin and C-reactive protein determination in diagnosed clinical samples

Early diagnosis of sepsis, combining blood cultures and inflammation biomarkers, continues to be a challenge, especially in very low birth weight (VLBW) infants because of limited availability of blood samples. Traditional diagnostic procedures are cumbersome, not fast enough, and require relatively large volumes of sample. Empiric use of antibiotics, before diagnostic confirmation, is required to decrease mortality, leading to potential antibiotic resistance and side effects in VLBW infants. To solve such a serious problem, a dual magnetoimmunosensor is proposed for simultaneous assessment of two of the most important sepsis biomarkers: procalcitonin (PCT for early phase) and Creactive protein (CRP for late phase). This ?sample-to-result? approach exhibited excellent sensitivity, selectivity, precision, and stability using low sample volumes (<30 ?L) and under 20 min of total assay. The analytical usefulness of the approach was demonstrated by analyzing clinically relevant samples of preterm neonates with suspicion of sepsis.Ministerio de Economía y CompetitividadComunidad Autónoma de MadridFundación “La Caixa

OFF-ON on-the-fly aptassay for rapid and accurate determination of procalcitonin in very low birth weight infants with sepsis suspicion

A fast and reliable OFF-ON micromotors (MM)-based aptassay for rapid, sensitive, and accurate determination of procalcitonin (PCT) as one of the leading neonatal sepsis biomarkers in very low birth weight infants? serum samples is proposed. MM were composed of three specific functional layers: graphene oxide (GO) as sensing layer, Ni for magnetic guidance and platinum nanoparticles (PtNPs) as catalytic layer for self-propulsion (MMGO/ Ni/PtNPs). The OFF-ON strategy relies on the binding of the aptamer tagged with alexa-405 as fluorescent label (AptPCT, ?em=447 nm) onto the GO outer layer of the MM (GOMM) to induce its fluorescence quenching (GOMMAptPCT, OFF state); which is next recovered due to selectively binding of the aptamer to the PCT and their separation from the GOMM surface (PCT-AptPCT, ON state). The fast OFF-ON on-the-fly aptassay allowed in just 5 min a highly selective and sensitive (LOD = 0.01 ng mL? 1) PCT determination in clinical samples from very low birth weight infants with sepsis suspicion, using only 25 ?L of sample in a clinically relevant range of concentrations (0.03?1280 ng mL? 1). The high concordance between the PCT levels obtained by our MMGO/Ni/PtNPsbased aptassay and those obtained by the Hospital, demonstrated the analytical potency of our approach for the analysis of neonatal samples as well as its potential to be used as a pre-diagnostic tool for sepsis.Ministerio de Ciencia e InnovaciónComunidad de MadridEuropean Commissio

Failure of early non-invasive ventilation in preterm infants with respiratory distress syndrome in current care practice in Spanish level-III neonatal intensive care units – a prospective observational study

Non-invasive respiratory ventilation; Preterm infant; Respiratory distress syndromeVentilación respiratoria no invasiva; Bebé prematuro; Síndrome de dificultad respiratoriaVentilació respiratòria no invasiva; Nadó prematur; Síndrome de dificultat respiratòriaIntroduction: Despite advances in respiratory distress syndrome (RDS) management over the past decade, non-invasive ventilation (NIV) failure is frequent and associated with adverse outcomes. There are insufficient data on the failure of different NIV strategies currently used in clinical practice in preterm infants. Methods: This was a prospective, multicenter, observational study of very preterm infants [gestational age (GA) <32 weeks] admitted to the neonatal intensive care unit for RDS that required NIV from the first 30 min after birth. The primary outcome was the incidence of NIV failure, defined as the need for mechanical ventilation for <72 h of life. Secondary outcomes were risk factors associated with NIV failure and complication rates. Results: The study included 173 preterm infants with a median GA of 28 (IQR 27–30) weeks and a median birth weight of 1,100 (IQR 800–1,333) g. The incidence of NIV failure was 15.6%. In the multivariate analysis, lower GA (OR, 0.728; 95% CI, 0.576–0.920) independently increased the risk of NIV failure. Compared to NIV success, NIV failure was associated with higher rates of unfavorable outcomes, including pneumothorax, intraventricular hemorrhage, periventricular leukomalacia, pulmonary hemorrhage, and a combined outcome of moderate-to-severe bronchopulmonary dysplasia or death. Conclusion: NIV failure occurred in 15.6% of the preterm neonates and was associated with adverse outcomes. The use of LISA and newer NIV modalities most likely accounts for the reduced failure rate. Gestational age remains the best predictor of NIV failure and is more reliable than the fraction of inspired oxygen during the first hour of life.This study was supported by Chiesi España, S.A.U., Barcelona, Spain. Chiesi España and S.A.U. were not involved in the collection and interpretation of the data or in the writing of the manuscript

On the Move-Sensitive Fluorescent Aptassay on Board Catalytic Micromotors for the Determination of Interleukin-6 in Ultra-Low Serum Volumes for Neonatal Sepsis Diagnostics

A graphene oxide/nickel/platinum nanoparticle micromotor (MM)-based fluorescent aptassay is proposed to determine interleukin-6 (IL-6) in serum samples from low-birth-weight infants (gestational age of less than 32 weeks and birthweight below 1000 g) with sepsis suspicion. In this kind of patients, IL-6 has demonstrated good sensitivity and specificity for the diagnosis of sepsis, both for early and late onset sepsis. The approach was based on the adsorption of the aptamer for IL-6 tagged with 6-FAM as a fluorescent label (AptIL?6, ?em = 520 nm) on the graphene oxide external layer (MMGO?AptIL?6) inducing fluorescence quenching (OFF state) and a subsequent on-the-move affinity recognition of IL-6 from AptIL?6 (IL-6?AptIL?6 complex) recovering the fluorescence (ON state). An aptamer against IL-6 was selected and developed by the systematic evolution of ligands by exponential enrichment technology. This approach displayed a suitable linear range of 0.07?1000 pg mL?1 (r = 0.995) covering the cut-off and clinical practice levels, allowing direct determination without any dilution and simplifying the analysis as well as exhibiting an excellent sensitivity (LOD = 0.02 pg mL?1) in ultralow volumes of diagnostic clinical samples (2 ?L). A high agreement between IL-6 levels obtained from our MM-based approach and the method used by the Hospital was obtained (relative error < 3%). The MM-based aptassay is competitive in comparison with that of the Hospital, in terms of a significant reduction of the sample volume (15 times less) and enhanced sensitivity, employing similar analysis times. These results position MM technology with enough potential to achieve high sensitivities in low sample volumes, opening new avenues in diagnosis based on low sample volumes.Ministerio de Ciencia e InnovaciónComunidad de MadridEuropean Commissio

The relationship between emotional intelligence, self-determined motivation and performance in canoeists

The results of recent studies indicate that athletes differ in the degree in which they perceive, process and regulate their emotions. The present study aims to utilize these findings in order to examine if self-determined motivation in sport could explain individual variations in emotional intelligence (EI) and results of sport competitions. A model of structural equations has proven these relationships in 386 canoeists from 35 countries. These results support the mediating factor of EI in relation to autonomous motivation and performance index (PI). The stability of EI and gender differences were analysed as a secondary objective. The results showed that EI stays relatively stable independent from the years of practice. However, rather surprisingly, men scored higher than women in emotional control and regulation as well as in empathy. These findings contribute to the study of EI in sport and have important implications for human performance in sport competition at high level.University Grants for the mobility and publication of results UPV/EHU (Cod.2254/2013)

Underdosing of Surfactant for Preterm Babies with Respiratory Distress Syndrome in Clinical Practice : A Retrospective Cohort Study

This study was supported by Chiesi España, S.A.U., Barcelona, Spain.Objective: To evaluate the initial doses of surfactant administered to preterm infants with respiratory distress syndrome. Study Design: This is a retrospective cohort study of 206 preterm infants admitted in four level III neonatal intensive care units of acute tertiary care hospitals in Spain between 2013 and 2015. Results: The mean initial dose of surfactant was 173.9 (37.3) mg/kg, and 47.5% of infants received a dose of 200 mg/kg ± 10% (180-220 mg/kg), 47% less than 180 mg/kg (-10%), and 5.4% more than 220 mg/kg (+10%). Very preterm infants (32 weeks). Conclusion: Administration of surfactant below the prescribed dose is a frequent error in clinical practice. Inadvertently rounding down doses seems a plausible explanation

The Impact of Postnatal Systemic Steroids on the Growth of Preterm Infants: A Multicenter Cohort Study

Postnatal steroids, often used to prevent and treat bronchopulmonary dysplasia, may influence the growth of preterm infants, although data are scarce in the literature. This is a multicenter cohort study including surviving preterm infants <32 weeks at birth (n = 17,621) from the Spanish Neonatal Network SEN1500 database, without major congenital malformations. Linear regression models were adjusted for postnatal steroids, respiratory severity course (invasive mechanical ventilation at 28 days), progression to moderate-severe bronchopulmonary dysplasia (O2 at 36 weeks), length of stay, sex, gestational age and z-scores at birth. A subgroup analysis depending on the timing of administration, ventilation status at 28 days and moderate-severe BPD diagnosis was also performed. Overall, systemic postnatal steroids were not independently associated with poorer weight gain (0.1; 95% CI: -0.05 to 0.2 g/kg/day), linear growth (0; 95% CI: -0.03 to 0.01 cm/week) or head circumference growth (-0.01; 95% CI: -0.02 to 0 cm/week). Patients who received steroids after 28 days or who were not O2 dependent at 36 weeks after having received steroids gained more weight (0.22; 95% CI: 0.04 to 0.4 and 0.2; 95% CI: 0.004 to 0.5 g/kg/day, respectively). Globally, systemic postnatal steroids had no significant adjusted effect on postnatal growth