A novel method for multiple cytokines/chemokines DropArray assay in 5 μL of bronchoalveolar lavage fluid (BALF) in mice

ObjectiveWe investigated the effects of chronic waterpipe (WP) smoke on pulmonary function and immune response in a murine model using a research-grade WP and the effects of acute exposure on the regulation of immediate-early genes (IEGs).MethodsWP smoke was generated using three WP smoke puffing regimens based on the Beirut regimen. WP smoke samples generated under these puffing regimens were quantified for nicotine concentration. Mice were chronically exposed for 6 months followed by assessment of pulmonary function and airway inflammation. Transcriptomic analysis using RNAseq was conducted after acute exposure to characterise the IEG response. These biomarkers were then compared with those generated after exposure to dry smoke (without water added to the WP bowl).ResultsWe determined that nicotine composition in WP smoke ranged from 0.4 to 2.5 mg per puffing session. The lung immune response was sensitive to the incremental severity of chronic exposure, with modest decreases in airway inflammatory cells and chemokine levels compared with air-exposed controls. Pulmonary function was unmodified by chronic WP exposure. Acute WP exposure was found to activate the immune response and identified known and novel IEG as potential biomarkers of WP exposure.ConclusionChronic exposure to WP smoke leads to immune suppression without significant changes to pulmonary function. Transcriptomic analysis of the lung after acute exposure to WP smoke showed activation of the immune response and revealed IEGs that are common to WP and dry smoke, as well as pools of IEGs unique to each exposure, identifying potential biomarkers specific to WP exposure.

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“…Samples were randomly positioned on a 96-well plate. The Curiox (Curiox Biosystems) wash system was used on plates, as reported previously 38 39…”

Section: Methodsmentioning

confidence: 99%

Immune modulation by chronic exposure to waterpipe smoke and immediate-early gene regulation in murine lungs

et al. 2019

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“…AA is found mainly in vegetarian foods and potato chips, which contain the highest levels of AA. Bread also contains high levels of AA ( 23 ). In this study, we have evaluated the effect of AA toxicity in an animal model of allergic asthma.…”

Section: Discussionmentioning

confidence: 99%

“…Also, one of the OVA-sensitized groups received AA by oral administration of AA-containing water. One other asthmatic group received a normal diet (without AA) (asthmatic control group) ( 23 ). In addition, one of the two healthy non-asthmatic groups was gavage-fed with the AA-containing water daily (in a similar way to asthmatic group that received AA).…”

Section: Methodsmentioning

confidence: 99%

Oral Administration of Acrylamide Worsens the Inflammatory Responses in the Airways of Asthmatic Mice Through Agitation of Oxidative Stress in the Lungs

et al. 2020

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Acrylamide is a toxic chemical substance produced when starch-rich foods are fried at high temperatures. Asthma is a chronic and complicated respiratory disease, of which genetic and environmental factors are the main triggers. Orally-received components may have an effect on asthma pathophysiology. The aim of this study was to investigate the role of AA as a stimulus in asthma. BALB/c mice were allocated into four groups as follows: two OVA-sensitized asthmatic groups, including one treated with AA by gavage feeding and one non-treated (asthma group), and two healthy (non-asthmatic) groups, one treated with AA by gavage feeding and one non-treated (negative control group). Airway hyperresponsiveness, cell count, cytokine levels in BAL fluid, lung histopathology, IgE levels, and oxidative stress indices including plasma level of MDA, pulmonary antioxidant enzymes (SOD and CAT) levels, HP content, and collagen fiber accumulation in lung tissue were measured. We found that the group of mice treated with both OVA and AA (asthmatic and AA-treated mice) experienced higher levels of asthma-associated biomarkers, including higher enhanced pause (Penh value), eosinophilic inflammation, mucus hyper secretion, goblet cell hyperplasia, total and OVA-specific IgE levels, IL-4, IL-5, and IL-13 levels than the group sensitized only with OVA (asthmatic mice). The OVA-AA-treated mice also experienced worsened levels of oxidative stress indicators. Healthy (non-asthmatic) mice that only received AA were in similar conditions to healthy untreated mice (negative control group). The OVA-AA-treated group showed more severe allergic asthma symptoms in comparison to the group only sensitized with OVA. Therefore, food/water contaminated with AA can act as a stimulant of allergic asthma and exacerbate the bronchial inflammatory responses.

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Magnetically aggregated biosensors for sensitive detection of biomarkers at low concentrations

Burg

Cohen

Margulis

et al. 2019

Applied Physics Letters

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Detection of biomarkers at low concentrations is essential for early diagnosis of numerous diseases. In many sensitive assays, the target molecules are tagged using fluorescently labeled probes and captured using magnetic beads. Magnetic beads facilitate washing and separation steps, are well suited for automation, and improve the assay sensitivity. Current devices rely on quantifying the target molecules by detecting the fluorescence signal from individual beads. Thus, to detect low concentrations of target molecules, these devices require sophisticated optical detectors, making them bulky and expensive. Here, we propose a compact fluorescence-based system that simply uses a small permanent magnet with a conic tip to aggregate the magnetic beads, forming a cluster of fluorescently labeled probes whose fluorescence signal is much greater than that of a single bead. Using the magnetically aggregated biosensors to detect human Interleukin-8, we demonstrated a limit of detection of 0.1 ng/l and a 4-log dynamic range performance, which is on par with the most sensitive devices but is achieved without their bulk and cost.

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A novel method for multiple cytokines/chemokines DropArray assay in 5 μL of bronchoalveolar lavage fluid (BALF) in mice

Cited by 4 publications

References 25 publications

Immune modulation by chronic exposure to waterpipe smoke and immediate-early gene regulation in murine lungs

Immune modulation by chronic exposure to waterpipe smoke and immediate-early gene regulation in murine lungs

Oral Administration of Acrylamide Worsens the Inflammatory Responses in the Airways of Asthmatic Mice Through Agitation of Oxidative Stress in the Lungs

Magnetically aggregated biosensors for sensitive detection of biomarkers at low concentrations

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