Continuous Flow vs. Static Chamber μPCR Devices on Flexible Polymeric Substrates

This paper was presented at the 4th Micro and Nano Flows Conference (MNF2014), which was held at University College, London, UK. The conference was organised by Brunel University and supported by the Italian Union of Thermofluiddynamics, IPEM, the Process Intensification Network, the Institution of Mechanical Engineers, the Heat Transfer Society, HEXAG - the Heat Exchange Action Group, and the Energy Institute, ASME Press, LCN London Centre for Nanotechnology, UCL University College London, UCL Engineering, the International NanoScience Community, www.nanopaprika.eu.Two types of μPCR devices, a continuous flow and a static chamber device, fabricated on flexible polymeric substrates are compared in the current computational study. Laminar flow, heat transfer in both solid and fluid, mass conservation of species, and reaction kinetics of PCR are coupled using COMSOL. The comparison is performed under same conditions; same material stack (based on flexible polymeric films with integrated microheaters), same species initial concentrations, amplification of the same volume of fluid sample, and implementation of the same PCR protocol. Performance is quantified in terms of DNA amplification, energy consumption, and total operating time. The calculations show that the efficiency of DNA amplification is higher in the continuous flow device. However, the continuous flow device requires (~6 times) greater energy consumption which is justified by the smaller thermal mass of the static chamber device. As regards the speed, the total time required for the static chamber μPCR is comparable to the time for the continuous flow μPCR

Achromatic rapid flares in hard X-rays in the γ\gamma-ray binary LS I +61-303

We report on the presence of very rapid hard X-ray variability in the $\gamma$-ray binary LS I +61 303. The results were obtained by analysing NuSTAR data, which show two achromatic strong flares on ks time-scales before apastron. The Swift-BAT orbital X-ray light curve is also presented, and the NuSTAR data are put in the context of the system orbit. The spectrum and estimated physical conditions of the emitting region indicate that the radiation is synchrotron emission from relativistic electrons, likely produced in a shocked pulsar wind. The achromaticity suggests that losses are dominated by escape or adiabatic cooling in a relativistic flow, and the overall behaviour in hard X-rays can be explained by abrupt changes in the size of the emitting region and/or its motion relative to the line of sight, with Doppler boosting potentially being a prominent effect. The rapid changes of the emitter could be the result of different situations such as quick changes in the intra-binary shock, variations in the re-accelerated shocked pulsar wind outside the binary, or strong fluctuations in the location and size of the Coriolis shock region. Although future multi-wavelength observations are needed to further constrain the physical properties of the high-energy emitter, this work already provides important insight into the complex dynamics and radiation processes in LS I +61-303.Comment: 8 pages, 7 figures. Accepted for publication in MNRA

A Passive Micromixer for Bioanalytical Applications

This paper was presented at the 4th Micro and Nano Flows Conference (MNF2014), which was held at University College, London, UK. The conference was organised by Brunel University and supported by the Italian Union of Thermofluiddynamics, IPEM, the Process Intensification Network, the Institution of Mechanical Engineers, the Heat Transfer Society, HEXAG - the Heat Exchange Action Group, and the Energy Institute, ASME Press, LCN London Centre for Nanotechnology, UCL University College London, UCL Engineering, the International NanoScience Community, www.nanopaprika.eu.Three passive micromixers with different geometries, i.e. zigzag, spiral, and split and merge (SaM) with labyrinthine channels, are compared with respect to their mixing efficiency by means of a computational study. The specifications are imposed from flexible printed circuit (FPC) technology which is used for their fabrication and from the applications to be implemented, i.e. the mixing of biochemical reagents. The computations include the numerical solution of continuity, Navier-Stokes, and mass conservation equations in 3d by ANSYS Fluent. The highest mixing efficiency is calculated for the SaM micromixer with the labyrinthine channel. Compared to a linear micromixer, the spiral micromixer improves the mixing efficiency by 8%, the zigzag by 11%, and the SaM by 92%; the diffusion coefficient of the biomolecule is 10-10 m2/s, the Reynolds number is 0.5, and the volume of each micromixer is 2.54 μl. The best of the three designs is realized by FPC technology and is experimentally evaluated by fluorescence microscopy

Effects of Weight-Bearing on Tibiofemoral, Patellofemoral, and Patellar Tendon Kinematics in Older Adults

Quantification of natural knee kinematics is essential for the assessment of joint function in the diagnosis of pathologies. Combined measurements of tibiofemoral and patellofemoral joint kinematics are necessary because knee pathologies, such as progression of osteoarthritis and patellar instability, are a frequent concern in both articulations. Combined measurement of tibiofemoral and patellofemoral kinematics also enables calculation of important quantities, specifically patellar tendon angle, which partly determines the loading vector at the tibiofemoral joint and patellar tendon moment arm. The goals of this research were to measure the differences in tibiofemoral and patellofemoral kinematics, patellar tendon angle (PTA), and patellar tendon moment arm (PTMA) that occur during non-weight-bearing and weight-bearing activities in older adults. Methods: High-speed stereo radiography was used to measure the kinematics of the tibiofemoral and patellofemoral joints in subjects as they performed seated, non-weight-bearing knee extension and two weight-bearing activities: lunge and chair rise. PTA and PTMA were extracted from the subject’s patellofemoral and tibiofemoral kinematics. Kinematics and the root mean square difference (RMSD) between non-weight-bearing and weight-bearing activities were compared across subjects and activities. Results: Internal rotation increased with weight-bearing (mean RMSD from knee extension was 4.2 ± 2.4° for lunge and 3.6 ± 1.8° for chair rise), and anterior translation was also greater (mean RMSD from knee extension was 2.2 ± 1.2 mm for lunge and 2.3 ± 1.4 mm for chair rise). Patellar tilt and medial–lateral translation changed from non-weight-bearing to weight-bearing. Changes of the patellar tendon from non-weight-bearing to weight-bearing were significant only for PTMA. Conclusions: While weight-bearing elicited changes in knee kinematics, in most degrees of freedoms, these differences were exceeded by intersubject differences. These results provide comparative kinematics for the evaluation of knee pathology and treatment in older adults

Molecular Characterization and Phylogenetic Analysis of Pseudomonas aeruginosa Isolates Recovered from Greek Aquatic Habitats Implementing the Double-Locus Sequence Typing Scheme.

The recently described double-locus sequence typing (DLST) scheme implemented to deeply characterize the genetic profiles of 52 resistant environmental Pseudomonas aeruginosa isolates deriving from aquatic habitats of Greece. DLST scheme was able not only to assign an already known allelic profile to the majority of the isolates but also to recognize two new ones (ms217-190, ms217-191) with high discriminatory power. A third locus (oprD) was also used for the molecular typing, which has been found to be fundamental for the phylogenetic analysis of environmental isolates given the resulted increased discrimination between the isolates. Additionally, the circulation of acquired resistant mechanisms in the aquatic habitats according to their genetic profiles was proved to be more extent. Hereby, we suggest that the combination of the DLST to oprD typing can discriminate phenotypically and genetically related environmental P. aeruginosa isolates providing reliable phylogenetic analysis at a local level

Prevalence and clinical implications of respiratory viruses in stable chronic obstructive pulmonary disease (COPD) and exacerbations: a systematic review and meta-analysis protocol.

INTRODUCTION: Both stable chronic obstructive pulmonary disease (COPD) and acute exacerbations represent leading causes of death, disability and healthcare expenditure. They are complex, heterogeneous and their mechanisms are poorly understood. The role of respiratory viruses has been studied extensively but is still not adequately addressed clinically. Through a rigorous evidence update, we aim to define the prevalence and clinical burden of the different respiratory viruses in stable COPD and exacerbations, and to investigate whether viral load of usual respiratory viruses could be used for diagnosis of exacerbations triggered by viruses, which are currently not diagnosed or treated aetiologically. METHODS AND ANALYSIS: Based on a prospectively registered protocol, we will systematically review the literature using standard methods recommended by the Cochrane Collaboration and the Grading of Recommendations Assessment, Development and Evaluation working group. We will search Medline/PubMed, Excerpta Medica dataBASE (EMBASE), the Cochrane Library, the WHO's Clinical Trials Registry and the proceedings of relevant international conferences on 2 March 2020. We will evaluate: (A) the prevalence of respiratory viruses in stable COPD and exacerbations, (B) differences in the viral loads of respiratory viruses in stable COPD vs exacerbations, to explore whether the viral load of prevalent respiratory viruses could be used as a diagnostic biomarker for exacerbations triggered by viruses and (C) the association between the presence of respiratory viruses and clinical outcomes in stable COPD and in exacerbations. ETHICS AND DISSEMINATION: Ethics approval is not required since no primary data will be collected. Our findings will be presented in national and international scientific conferences and will be published in peer reviewed journals. Respiratory viruses currently represent a lost opportunity to improve the outcomes of both stable COPD and exacerbations. Our work aspires to 'demystify' the prevalence and clinical burden of viruses in stable COPD and exacerbations and to promote clinical and translational research. PROSPERO REGISTRATION NUMBER: CRD42019147658

Diagnostic importance of pulmonary interleukin-1beta and interleukin-8 in ventilator-associated pneumonia.

BACKGROUND: Ventilator-associated pneumonia (VAP) is the most commonly fatal nosocomial infection. Clinical diagnosis of VAP remains notoriously inaccurate. The hypothesis was tested that significantly augmented inflammatory markers distinguish VAP from conditions closely mimicking VAP. METHODS: A prospective, observational cohort study was carried out in two university hospital intensive care units recruiting 73 patients with clinically suspected VAP, and a semi-urban primary care practice recruiting a reference group of 21 age- and sex-matched volunteers. Growth of pathogens at >10(4) colony-forming units (cfu)/ml of bronchoalveolar lavage fluid (BALF) distinguished VAP from "non-VAP". Inflammatory mediators were quantified in BALF and serum. Mediators showing significant differences between patients with and without VAP were analysed for diagnostic utility by receiver operator characteristic (ROC) curves. RESULTS: Seventy-two patients had recoverable lavage-24% had VAP. BALF interleukin-1beta (IL-1beta), IL-8, granulocyte colony-stimulating factor and macrophage inflammatory protein-1alpha were significantly higher in the VAP group (all p<0.005). Using a cut-off of 10 pg/ml, BALF IL-1beta generated negative likelihood ratios for VAP of 0.09. In patients with BALF IL-1beta <10 pg/ml the post-test probability of VAP was 2.8%. Using a cut-off value for IL-8 of 2 ng/ml, the positive likelihood ratio was 5.03. There was no difference in cytokine levels between patients with sterile BALF and those with growth of <10(4) cfu/ml. CONCLUSIONS: BALF IL-1beta and IL-8 are amongst the strongest markers yet identified for accurately demarcating VAP within the larger population of patients with suspected VAP. These findings have potential implications for reduction in unnecessary antibiotic use but require further validation in larger populations

Diagnostic accuracy of pulmonary host inflammatory mediators in the exclusion of ventilator-acquired pneumonia.

BACKGROUND: Excessive use of empirical antibiotics is common in critically ill patients. Rapid biomarker-based exclusion of infection may improve antibiotic stewardship in ventilator-acquired pneumonia (VAP). However, successful validation of the usefulness of potential markers in this setting is exceptionally rare. OBJECTIVES: We sought to validate the capacity for specific host inflammatory mediators to exclude pneumonia in patients with suspected VAP. METHODS: A prospective, multicentre, validation study of patients with suspected VAP was conducted in 12 intensive care units. VAP was confirmed following bronchoscopy by culture of a potential pathogen in bronchoalveolar lavage fluid (BALF) at >10(4) colony forming units per millilitre (cfu/mL). Interleukin-1 beta (IL-1β), IL-8, matrix metalloproteinase-8 (MMP-8), MMP-9 and human neutrophil elastase (HNE) were quantified in BALF. Diagnostic utility was determined for biomarkers individually and in combination. RESULTS: Paired BALF culture and biomarker results were available for 150 patients. 53 patients (35%) had VAP and 97 (65%) patients formed the non-VAP group. All biomarkers were significantly higher in the VAP group (p<0.001). The area under the receiver operator characteristic curve for IL-1β was 0.81; IL-8, 0.74; MMP-8, 0.76; MMP-9, 0.79 and HNE, 0.78. A combination of IL-1β and IL-8, at the optimal cut-point, excluded VAP with a sensitivity of 100%, a specificity of 44.3% and a post-test probability of 0% (95% CI 0% to 9.2%). CONCLUSIONS: Low BALF IL-1β in combination with IL-8 confidently excludes VAP and could form a rapid biomarker-based rule-out test, with the potential to improve antibiotic stewardship