Mediating Role of Glucose-Lipid Metabolism in the Association between the Increased Risk of Coronary Heart Disease and Exposure to Organophosphate Esters,Phthalates,and Polycyclic Aromatic Hydrocarbons

There is a lack of human evidence concerning the cardiovascular effects of combined exposure to endocrine disruptors.This case-control study sought to investigate coronary heart disease(CHD)associations with exposure to organophosphate flame retardants(OFRs),phthalates(PAEs),and polycyclic aromatic hydrocarbons(PAHs)among 148 adults with coronary-angiography-diagnosed CHD and 320 healthy adults from southern China.The mediating role of glucose-lipid metabolism was also explored.Bayesian kernel machine regression suggested that when exposure status was fixed to the 75th percentile with the median value as the reference,exposure to OFRs,PAEs,and PAHs was associated with an 84%(95%CI:36%−134%),132%(12%−252%),and 214%(89%−331%)increased risk of developing CHD,respectively.Weighted quantile sum regression indicated urinary bis(2-butoxyethyl)phosphate(BBOEP),dibutyl phosphate(DBP),monoisononyl phthalate(miNP),and metabolites of phenanthrene may be major contributors to the overall effect of mixtures.In further analyses on identified chemical risk factors,mediation analyses suggested exposure to phenanthrene may increase the risk of CHD via elevating total cholesterol and blood glucose,while exposure to DiNP mainly associates with serum lipids.Besides,we observed a slight mediation effect of oxidative DNA damage between urinary BBOEP and risk of CHD.These results provide potential direction for further experimental studies.Longitudinal evidence is needed to clarify the causation of the results.

On-demand light-driven release of droplets stabilized via a photoresponsive fluorosurfactant

Water-in-oil droplets have emerged as promising microreactors for high-throughput biochemical analysis due to their features of reduced sample consumption and automated operation.For a typical screening application,droplets are often trapped for continuous monitoring of the reaction over an extended period,followed by the selective retrieval of targeted droplets based on the after-effect of biochemical reactions.While techniques for droplet trapping are well developed,retrieval of targeted droplets mainly demands complicated device fabrication or sophisticated control.Herein,facile and rapid selective droplet release is achieved by utilizing a new class of photoresponsive fluorosurfactant based on plasmonic nanoparticles.The intense photothermal response provided by this novel photoresponsive fluorosurfactant is capable of vaporizing the fluorocarbon oil at the droplet interface under laser illumination,resulting in a bubble releasing a trapped droplet on demand.A fully automated fluorescence-activated droplet release platform has also been developed to demonstrate its potential for droplet-based large-scale screening applications.

Double emulsion-pretreated microwell culture for the in vitro production of multicellular spheroids and their in situ analysis

Multicellular spheroids have served as a promising preclinical model for drug efficacy testing and disease modeling.Many microfluidic tech no logies,in eluding those based on water-oil-water double emulsions,have been introduced for the production of spheroids.However,sustained culture and the in situ characterization of the gen erated spheroids are currently unavailable for the double emulsion-based spheroid model.This study presents a streamlined workflow,termed the double emulsion-pretreated microwell culture(DEPMiC),incorporating the features of(1)effective initiation of uniform-sized multicellular spheroids by the pretreatment of double emulsions produced by microfluidics without the requirement of biomaterial scaffolds;(2)sustained maintenance and culture of the produced spheroids with facile removal of the oil confinement;and(3)in situ characterization of individual spheroids localized in microwells by a built-in analytical station.Characterized by microscopic observations and Raman spectroscopy,the DEPMiC cultivated spheroids accumulated elevated lipid ordering on the apical membran巳similar to that observed in their Matrigel counterparts.Made possible by the proposed tech no logical advancement,this study subsequently examined the drug responses of these in vitro-generated multicellular spheroids.The developed DEPMiC platform is expected to gen erate health benefits in personalized cancer treatment by offering a pre-animal tool to dissect heterogeneity from individual tumor spheroids.

Demarcating the membrane damage for the extraction of functional mitochondria

Defective mitochondria have been linked to several critical human diseases such as neurodegenerative disorders,cancers and cardiovascular disease.However,the detailed characterization of mitochondria has remained relatively unexplored,largely due to the lack of effective extraction methods that may sufficiently retain the functionality of mitochondria,particularly when limited amount of sample is considered.In this study,we explore the possibility of modulating hydrodynamic stress through a cross-junction geometry at microscale to selectively disrupt the cellular membrane while mitochondrial membrane is secured.The operational conditions are empirically optimized to effectively shred the cell membranes while keeping mitochondria intact for the model mammalian cell lines,namely human embryonic kidney cells,mouse muscle cells and neuroblastoma cells.Unsurprisingly,the disruption of cell membranes with higher elastic moduli(neuroblastoma)requires elevated stress.This study also presents a comparative analysis of total protein yield and concentrations of extracted functional mitochondria with two commercially available mitochondria extraction approaches,the Dounce Homogenizer and the Qproteome®Mitochondria Isolation Kit,in a range of cell concentrations.Our findings show that the proposed“microscale cell shredder”yields at least 40%more functional mitochondria than the two other approaches and is able to preserve the morphological integrity of extracted mitochondria,particularly at low cell concentrations(5–20×10^(4) cells/mL).Characterized by its capability of rapidly processing a limited quantity of samples(200μL),demarcating the membrane damage through the proposed microscale cell shredder represents a novel strategy to extract subcellular organelles from clinical samples.