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1.
J Vis Exp ; (140)2018 10 20.
Article in English | MEDLINE | ID: mdl-30394380

ABSTRACT

A significant number of lead compounds fail in the pharmaceutical pipeline because animal studies often fail to predict clinical responses in human patients. Human Organ-on-a-Chip (Organ Chip) microfluidic cell culture devices, which provide an experimental in vitro platform to assess efficacy, toxicity, and pharmacokinetic (PK) profiles in humans, may be better predictors of therapeutic efficacy and safety in the clinic compared to animal studies. These devices may be used to model the function of virtually any organ type and can be fluidically linked through common endothelium-lined microchannels to perform in vitro studies on human organ-level and whole body-level physiology without having to conduct experiments on people. These Organ Chips consist of two perfused microfluidic channels separated by a permeable elastomeric membrane with organ-specific parenchymal cells on one side and microvascular endothelium on the other, which can be cyclically stretched to provide organ-specific mechanical cues (e.g., breathing motions in lung). This protocol details the fabrication of flexible, dual channel, Organ Chips through casting of parts using 3D printed molds, enabling combination of multiple casting and post-processing steps. Porous poly (dimethyl siloxane) (PDMS) membranes are cast with micrometer sized through-holes using silicon pillar arrays under compression. Fabrication and assembly of Organ Chips involves equipment and steps that can be implemented outside of a traditional cleanroom. This protocol provides researchers with access to Organ Chip technology for in vitro organ- and body-level studies in drug discovery, safety and efficacy testing, as well as mechanistic studies of fundamental biological processes.


Subject(s)
Cell Culture Techniques/instrumentation , Microfluidics/methods , Animals , Humans
2.
Appl Spectrosc ; 69(7): 785-93, 2015 Jul.
Article in English | MEDLINE | ID: mdl-26036870

ABSTRACT

A protocol created for acephate detection on particulates and vapors surrounding farmworkers as well as in urine samples is reported. Acephate is detected to the low parts-per-billion (ppb) range using surface-enhanced Raman spectroscopy (SERS). Optimal SERS sensor metal choice and post-production treatments to improve sensor stability in aqueous solutions containing acephate are presented. Acephate is detected in the vapor phase and can be differentiated from urine components and structurally similar pesticides, including the acephate metabolite-degradation product methamidophos. Protocol evaluation and preliminary field tests from North Carolina farms are discussed.


Subject(s)
Organothiophosphorus Compounds/urine , Pesticides/urine , Phosphoramides/urine , Spectrum Analysis, Raman/methods , Humans , Occupational Exposure/analysis , Organothiophosphorus Compounds/analysis , Pesticides/analysis , Urinalysis/methods , Volatilization , Water/analysis
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