Inter-Establishment Complex Musculoskeletal Care Pathways in Montreal: Timeline of a Collaboration Involving a Research Team Within a Continuous Quality Improvement Initiative.

Children and adolescents with complex musculoskeletal conditions may receive health care that requires at least 1 transfer between 4 specialized pediatric establishments in the Montreal region (Québec, Canada). This may result in challenges in navigating the system. A collaborative approach, aiming to make the inter-establishment care pathways seamless and to improve the integration of musculoskeletal health services, brought together key stakeholders including a research team. The aim of this paper is to describe the timeline of the collaborative approach's key milestones and activities and, more specifically, to describe the context, process, and outputs of the involvement of researchers in support of a continuous quality improvement project based on an integrated approach. The descriptive timeline was constructed from a qualitative document analysis of the project-related gray literature (n = 80 documents) and was validated and interpreted with key stakeholders. The results showed how the collaborative project was set up and operated, as well as what solutions were developed and implemented. The strategies on how the research team was involved in the integrated approach in addition to its research activities were also described. Conclusions suggest practice recommendations for creating change processes by integrating research, service evaluation and clinical audit into quality improvement projects.

Droplet-based microfluidics platform for antifungal analysis against filamentous fungi.

Fungicides are extensively used in agriculture to control fungal pathogens which are responsible for significant economic impact on plant yield and quality. The conventional antifungal screening techniques, such as water agar and 96-well plates, are based on laborious protocols and bulk analysis, restricting the analysis at the single spore level and are time consuming. In this study, we present a droplet-based microfluidic platform that enables antifungal analysis of single spores of filamentous fungus Alternaria alternata. A droplet-based viability assay was developed, allowing the germination and hyphal growth of single A. alternata spores within droplets. The viability was demonstrated over a period of 24 h and the antifungal screening was achieved using Kunshi/Tezuma as antifungal agent. The efficacy results of the droplet-based antifungal analysis were compared and validated with the results obtained from conventional protocols. The percentage inhibitions assessed by the droplet-based platform were equivalent with those obtained by the other two methods, and the Pearson correlation analysis showed high correlation between the three assays. Taken together, this droplet-based microfluidic platform provides a wide range of potential applications for the analysis of fungicide resistance development as well as combinatorial screening of other antimicrobial agents and even antagonistic fungi.

Bacterial Expression Systems for Enzymatic Activity in Droplet-Based Microfluidics.

Functional screenings in droplet-based microfluidics require the analysis of various types of activities of individual cells. When screening for enzymatic activities, the link between the enzyme of interest and the information-baring molecule, the DNA, must be maintained to relate phenotypes to genotypes. This linkage is crucial in directed evolution experiments or for the screening of natural diversity. Micro-organisms are classically used to express enzymes from nucleic acid sequences. However, little information is available regarding the most suitable expression system for the sensitive detection of enzymatic activity at the single-cell level in droplet-based microfluidics. Here, we compare three different expression systems for l-asparaginase (l-asparagine amidohydrolase, EC 3.5.1.1), an enzyme of therapeutic interest that catalyzes the conversion of l-asparagine to l-aspartic acid and ammonia. We developed three expression vectors to produce and localize l-asparaginase (l-ASNase) in E. coli either in the cytoplasm, on the surface of the inner membrane (display), or in the periplasm. We show that the periplasmic expression is the most optimal strategy combining both a good yield and a good accessibility for the substrate without the need for lysing the cells. We suggest that periplasmic expression may provide a very efficient platform for screening applications at the single-cell level in microfluidics.

High-Throughput Triggered Merging of Surfactant-Stabilized Droplet Pairs Using Traveling Surface Acoustic Waves.

We present an acoustofluidic device for fluorescently triggered merging of surfactant-stabilized picoliter droplet pairs at high throughput. Droplets that exceed a preset fluorescence threshold level are selectively merged by a traveling surface acoustic wave (T-SAW) pulse. We characterize the operation of our device by analyzing the merging efficiency as a function of acoustic pulse position, duration, and acoustic pressure amplitude. We probe droplet merging at different droplet rates and find that efficient merging occurs above a critical acoustic power level. Our results indicate that the efficiency of acoustically induced merging of surfactant stabilized droplets is correlated with acoustic streaming velocity. Finally, we discuss how both time-averaged and instantaneous acoustic pressure fields can affect the integrity of surfactant layers. Our technique, by allowing the merging of up to 105 droplets per hour, shows great potential for integration into microfluidic systems for high-throughput and high-content screening applications.