Explore the vast range of titles available.

The most performing techniques enabling early diagnosis of infectious diseases rely on nucleic acid detection. Today, because of their high technicality and cost, nucleic acid amplification tests (NAAT) are of benefit only to a small fraction of developing countries population. By reducing costs, simplifying procedures and enabling multiplexing, paper microfluidics has the potential to considerably facilitate their accessibility. However, most of the studies performed in this area have not quit the lab. This letter brings NAAT on paper closer to the field, by using clinical samples and operating in a resource-limited setting. We first performed isothermal reverse transcription and Recombinase Polymerase Amplification (RT-RPA) of synthetic Ribonucleic Acid (RNA) of Ebola virus using paper microfluidics devices. We further applied this method in Guinea to detect the presence of Ebola virus in human sample RNA extracts, with minimal facilities (carry-on detection device and freeze-dried reagents on paper). RT-RPA results were available in few minutes and demonstrate a sensitivity of 90.0% compared to the gold-standard RT-PCR on a set of 43 patient samples. Furthermore, the realization of a nine-spot multilayered device achieving the parallel detection of three distinct RNA sequences opens a route toward the detection of multiple viral strains or pathogens.

To respond to the urgent need for COVID-19 testing, countries perform nucleic acid amplification tests (NAAT) for the detection of SARS-CoV-2 in centralized laboratories. Real-time RT—PCR (Reverse transcription—Polymerase Chain Reaction), used to amplify and detect the viral RNA., is considered, as the current gold standard for diagnostics. It is an efficient process, but the complex engineering required for automated RNA extraction and temperature cycling makes it incompatible for use in point of care settings [1]. In the present work, by harnessing progress made in the past two decades in isothermal amplification and paper microfluidics, we created a portable test, in which SARS-CoV-2 RNA is extracted, amplified isothermally by RT—LAMP (Loop-mediated Isothermal Amplification), and detected using intercalating dyes or fluorescent probes. Depending on the viral load in the tested samples, the detection takes between twenty minutes and one hour. Using a set of 16 pools of naso-pharyngal swab eluates, we estimated a limit of detection comparable to real-time RT-PCR (i.e. 1 genome copies per microliter of clinical sample) and no cross‐reaction with eight major respiratory viruses currently circulating in Europe. We designed and fabricated an easy-to-use portable device called “COVIDISC” to carry out the test at the point of care. The low cost of the materials along with the absence of complex equipment will expedite the widespread dissemination of this device. What is proposed here is a new efficient tool to help managing the pandemics.

Aluminium salts such as aluminium chlorohydrate (ACH) are the active ingredients of antiperspirant products. Their mechanism of action involves a temporary and superficial plugging of eccrine sweat pores at the skin surface. We developed a microfluidic system that allows the real time observation of the interactions between sweat and ACH in conditions mimicking physiological sweat flow and pore dimensions. Using artificial sweat containing bovine serum albumin as a model protein, we performed experiments under flowing conditions to demonstrate that pore clogging results from the aggregation of proteins by aluminium polycations at specific location in the sweat pore. Combining microfluidic experiments, confocal microscopy and numerical models helps to better understand the physical chemistry and mechanisms involved in pore plugging. The results show that plugging starts from the walls of sweat pores before expanding into the centre of the channel. The simulations aid in explaining the influence of ACH concentration as well as the impact of flow conditions on the localization of the plug. Altogether, these results outline the potential of both microfluidic confocal observations and numerical simulations at the single sweat pore level to understand why aluminium polycations are so efficient for sweat channel plugging.

The concept of using stimuli-responsive hydrogels to actuate fluids in microfluidic devices is particularly attractive, but limitations, in terms of spatial resolution, speed, reliability and integration, have hindered its development during the past two decades. By patterning and grafting poly(N-isopropylacrylamide) PNIPAM hydrogel films on plane substrates with a 2 μm horizontal resolution and closing the system afterward, we have succeeded in unblocking bottlenecks that thermo-sensitive hydrogel technology has been challenged with until now. In this paper, we demonstrate, for the first time with this technology, devices with up to 7800 actuated micro-cages that sequester and release solutes, along with valves actuated individually with closing and opening switching times of 0.6±0.1 and 0.25±0.15 s, respectively. Two applications of this technology are illustrated in the domain of single cell handling and the nuclear acid amplification test (NAAT) for the Human Synaptojanin 1 gene, which is suspected to be involved in several neurodegenerative diseases such as Parkinson's disease. The performance of the temperature-responsive hydrogels we demonstrate here suggests that in association with their moderate costs, hydrogels may represent an alternative to the actuation or handling techniques currently used in microfluidics, that are, pressure actuated polydimethylsiloxane (PDMS) valves and droplets.

The goal of this work is to make progress in the domain of near-wall velocimetry. The technique we use is based on the tracking of nanoparticles in an evanescent field, close to a wall, a technique called TIRF (total internal reflection fluorescence)-based velocimetry. The particles are filmed continuously, with no time gap between two frames, so that no information on their trajectories is lost. A number of biases affect the measurements: Brownian motion, heterogeneities induced by the walls, statistical biases, photobleaching, polydispersivity and limited depth of field. Their impacts are quantified by carrying out Langevin stochastic simulations, in a way similar to Guasto & Breuer (Exp. Fluids, vol. 47, 2009, pp. 1059–1066). By using parameters calibrated separately or known, we obtain satisfactory agreement between experiments and simulations, concerning the intensity density distributions, velocity fluctuation distributions and the slopes of the linear velocity profiles. Slip lengths measurements, taken as benchmarks for analysing the performances of the technique, are carried out by extrapolating the corrected velocity profiles down to the origin along with determining the wall position with an unprecedented accuracy. For hydrophilic surfaces, we obtain $1\\pm 5~\\text{nm}$ for the slip length in sucrose solutions and $9\\pm 10~\\text{nm}$ in water, and for hydrophobic surfaces, $32\\pm 5~\\text{nm}$ for sucrose solutions and $55\\pm 9~\\text{nm}$ for water. The errors (based on 95 % confidence intervals) are significantly smaller than the state of the art, but more importantly, the method demonstrates for the first time a capacity to measure slippage with a satisfactory accuracy, while providing a local information on the flow structure with a nanometric spatial precision and velocity errors of a few per cent. Our study confirms the discrepancy already pointed out in the literature between numerical and experimental slip length estimates. With the progress conveyed by the present work, TIRF-based technique with continuous tracking can be considered as a quantitative method for investigating flow properties close to walls, providing both global and local information on the flow.

Dried blood spot (DBS) has risen in popularity due to the ease of sampling, storing, shipping and more. Despite those advantages, recovery of the dried blood in solution for analysis is still a bottleneck as it is manual, time-consuming and leads to high dilutions. To overcome those issues, we have developed a microfluidic chip allowing reversible opening, holding hermetically DBS and forcing the elution buffer through the thickness of DBS. The new technique, validated with clinical samples, is automated, fast, robust, precise, compatible with in-line analysis and leads to a highly concentrated extraction. Moreover, by using model experiments with fluorescein solutions, we show that the elution process is governed by an advection-diffusion coupling commonly known as Taylor dispersion. This new technique could open the way to a new generation of analytical devices to quantify analytes in DBS samples for a wide range of applications.

The goal of this work is to make progress in the domain of near-wall velocimetry. The technique we use is based on the tracking of nanoparticles in an evanescent field, close to a wall, a technique called TIRF (Total Internal Reflection Fluorescence)-based velocimetry. At variance with the methods developed in the literature, we permanently keep track of the light emitted by each particle during the time the measurements of their positions ('altitudes') and speeds are performed. By performing the Langevin simulation, we quantified effect of biases such as Brownian motion, heterogeneities induced by the walls, statistical biases, photo bleaching, polydispersivity and limited depth of field. Using this method, we obtained slip length on hydrophilic surfaces of 1\\( \\pm \\)5 nm for sucrose solution, and 9\\( \\pm \\)10 nm for water; On hydrophobic surface, 32\\( \\pm \\)5 nm for sucrose solution, and 55\\( \\pm \\)9 nm for water. The errors (based on 95% confidence intervals) are significantly smaller than the state-of-the-art, but more importantly, the method demonstrates for the first time a capacity to measure slippage with a satisfactory accuracy, while providing a local information on the flow structure with a nanometric resolution. Our study confirms the discrepancy already pointed out in the literature between numerical and experimental slip length estimates. With the progress conveyed by the present work, TIRF based technique with continuous tracking can be considered as a quantitative method for investigating flow properties close to walls, providing both global and local information on the flow.

We have postulated that the aryl hydrocarbon receptor (AHR) drives the later, more lethal stages of some cancers when chronically activated by endogenous ligands. However, other studies have suggested that, under some circumstances, the AHR can oppose tumor aggression. Resolving this apparent contradiction is critical to the design of AHR-targeted cancer therapeutics. Molecular (siRNA, shRNA, AHR repressor, CRISPR-Cas9) and pharmacological (AHR inhibitors) approaches were used to confirm the hypothesis that AHR inhibition reduces human cancer cell invasion (irregular colony growth in 3D Matrigel cultures and Boyden chambers), migration (scratch wound assay) and metastasis (human cancer cell xenografts in zebrafish). Furthermore, these assays were used for a head-to-head comparison between AHR antagonists and agonists. AHR inhibition or knockdown/knockout consistently reduced human ER−/PR−/Her2− and inflammatory breast cancer cell invasion, migration, and metastasis. This was associated with a decrease in invasion-associated genes (e.g., Fibronectin, VCAM1, Thrombospondin, MMP1) and an increase in CDH1/E-cadherin, previously associated with decreased tumor aggression. Paradoxically, AHR agonists (2,3,7,8-tetrachlorodibenzo-p-dioxin and/or 3,3′-diindolylmethane) similarly inhibited irregular colony formation in Matrigel and blocked metastasis in vivo but accelerated migration. These data demonstrate the complexity of modulating AHR activity in cancer while suggesting that AHR inhibitors, and, under some circumstances, AHR agonists, may be useful as cancer therapeutics.

Background Head and neck squamous cell carcinoma (HNSCC) is an aggressive malignancy characterized by tumor heterogeneity, locoregional metastases, and resistance to existing treatments. Although a number of genomic and molecular alterations associated with HNSCC have been identified, they have had limited impact on the clinical management of this disease. To date, few targeted therapies are available for HNSCC, and only a small fraction of patients have benefited from these treatments. A frequent feature of HNSCC is the inappropriate activation of β-catenin that has been implicated in cell survival and in the maintenance and expansion of stem cell-like populations, thought to be the underlying cause of tumor recurrence and resistance to treatment. However, the therapeutic value of targeting β-catenin activity in HNSCC has not been explored. Methods We utilized a combination of computational and experimental profiling approaches to examine the effects of blocking the interaction between β-catenin and cAMP-responsive element binding (CREB)-binding protein (CBP) using the small molecule inhibitor ICG-001. We generated and annotated in vitro treatment gene expression signatures of HNSCC cells, derived from human oral squamous cell carcinomas (OSCCs), using microarrays. We validated the anti-tumorigenic activity of ICG-001 in vivo using SCC-derived tumor xenografts in murine models, as well as embryonic zebrafish-based screens of sorted stem cell-like subpopulations. Additionally, ICG-001-inhibition signatures were overlaid with RNA-sequencing data from The Cancer Genome Atlas (TCGA) for human OSCCs to evaluate its association with tumor progression and prognosis. Results ICG-001 inhibited HNSCC cell proliferation and tumor growth in cellular and murine models, respectively, while promoting intercellular adhesion and loss of invasive phenotypes. Furthermore, ICG-001 preferentially targeted the ability of subpopulations of stem-like cells to establish metastatic tumors in zebrafish. Significantly, interrogation of the ICG-001 inhibition-associated gene expression signature in the TCGA OSCC human cohort indicated that the targeted β-catenin/CBP transcriptional activity tracked with tumor status, advanced tumor grade, and poor overall patient survival. Conclusions Collectively, our results identify β-catenin/CBP interaction as a novel target for anti-HNSCC therapy and provide evidence that derivatives of ICG-001 with enhanced inhibitory activity may serve as an effective strategy to interfere with aggressive features of HNSCC.

OBJECTIVE:--Recent studies suggested that the blockade of the renin-angiotensin system (RAS) may be associated with metabolic benefits. However, data about the potential influence of the ACE insertion/deletion (I/D) genotype on insulin resistance have been contradictory with studies of limited sample sizes. The purpose of this study was to investigate the relationship between the ACE gene I/D polymorphism and both insulin sensitivity and glucose intolerance in a large cohort of healthy subjects. RESEARCH DESIGN AND METHODS--A total of 1,286 participants in the Relationship Between Insulin Sensitivity and Cardiovascular Disease Risk Study had a 75-g oral glucose tolerance test and a hyperinsulinemic-euglycemic clamp to assess whole-body insulin sensitivity. RESULTS:--Age, BMI, waist, fat-free mass (ffm), and physical activity did not differ by ACE genotype. Fasting glucose and insulin were similar among genotypes, but 2-h glucose levels were higher in DD than in ID and II subjects (DD: 5.9 ± 1.7; ID: 5.7 ± 1.5; II: 5.6 ± 1.5 mmol/l) (P = 0.004). Participants with the DD genotype were more likely to have impaired glucose tolerance than those with the ID and II genotypes (13.1 vs. 8.7%; P = 0.02). Insulin sensitivity was lower in participants with the DD genotype than in those with the II genotype (136 ± 63 vs. 147 ± 65 μmol · min⁻¹· kg ffm⁻¹ · mmol⁻¹ · l⁻¹; P = 0.02). The presence of the D allele was associated with a trend, albeit not significant, for reduced insulin secretion during the oral glucose tolerance test (P = 0.07). CONCLUSIONS:--The ACE I/D polymorphism is associated with whole-body insulin sensitivity and with impaired glucose tolerance in our healthy population. These findings confirm potential interactions between the RAS and glucose metabolism.