Customizable Multiplex Antibody Array Immunoassays with Attomolar Sensitivities.

We have developed a customizable contact printed multiplex immunoassay capable of simultaneously measuring up to five analytes with attomolar sensitivities. This enzyme-linked immunosorbent assay (ELISA) was based on spotting different antibodies in a circular pattern at the bottom of a microtiter plate well. Unlike traditional antibody printing for ELISA that prints a capture antibody specific to a target of interest, in this ELISA we printed unique "anchor" antibodies at the well surface, each having a high affinity for a specific peptide target. By coupling each peptide to a unique assay capture antibody, this array of anchor antibodies enabled a customizable contact printed multiplex immunoassay workflow. As a proof of concept, we developed a 5-plex assay measuring interleukin 5 (IL-5), interleukin 6 (IL-6), interleukin 10 (IL-10), interleukin 22 (IL-22), and tumor necrosis factor alpha (TNF-α). Measurements of these five analytes in serum and plasma correlated well between the method utilizing the anchor antibodies and peptides and the traditional capture antibody printing approach, with r2 values of 0.99, 0.93, 0.99, 0.96, and 0.75 for IL-5, IL-6, IL-10, IL-22, and TNFα, respectively. This approach makes customizable multiplex ultrasensitive ELISA available to laboratories without access to the precision printing instrumentation and will be useful for antibody screening, custom assay development, biomarker detection, and protein profiling for diagnostic applications.

Nursing's Critical Role in the Shifting Landscape of Mental Health.

By 2020, mental and substance use disorders will surpass all physical diseases as a major cause of disability in the United States. Four key actions are proposed in which healthcare systems and nurses--the largest group of providers--can leverage nursing to address the biggest public health challenge the United States and many other nations face. Faith community nurses and faith congregations have particular opportunities to address this overwhelming need.

Laser-Ablated Vortex Fluidic-Mediated Synthesis of Superparamagnetic Magnetite Nanoparticles in Water Under Flow.

Selective formation of only one iron oxide phase is a major challenge in conventional laser ablation process, as is scaling up the process. Herein, superparamagnetic single-phase magnetite nanoparticles of hexagonal and spheroidal-shape, with an average size of ca. 15 nm, are generated by laser ablation of bulk iron metal at 1064 nm in a vortex fluidic device (VFD). This is a one-step continuous flow process, in air at ambient pressure, with in situ uptake of the nanoparticles in the dynamic thin film of water in the VFD. The process minimizes the generation of waste by avoiding the need for any chemicals or surfactants and avoids time-consuming purification steps in reducing any negative impact of the processing on the environment.

GPCR screening via ERK 1/2: a novel platform for screening G protein-coupled receptors.

Discovery of novel agonists and antagonists for G protein-coupled receptors (GPCRs) relies heavily on cell-based assays because determination of functional consequences of receptor engagement is often desirable. Currently, there are several key parameters measured to achieve this, including mobilization of intracellular Ca2+ and formation of cyclic adenosine monophosphate or inositol triphosphate. However, no single assay platform is suitable for all situations, and all of the assays have limitations. The authors have developed a new high-throughput homogeneous assay platform for GPCR discovery as an alternative to current assays, which employs detection of phosphorylation of the key signaling molecule p42/44 MAP kinase (ERK 1/2). The authors show that ERK 1/2 is consistently activated in cells stimulated by Gq-coupled GPCRs and provides a new high-throughput platform for screening GPCR drug candidates. The activation of ERK 1/2 in Gq-coupled GPCR systems generates comparable pharmacological data for receptor agonist and antagonist data obtained by other GPCR activation measurement techniques.

Development and validation of a single-well cell-based assay for the detection of endogenous phosphoproteins.

We describe a cellular assay for detection of phosphorylation of endogenous proteins, whereby cells are seeded, treated, and assayed for modulation of phosphorylation in a single microplate well. The procedure is coupled to a rapid, one-wash sandwich enzyme-linked immuno-sorbent assay, enabling results to be obtained within 3-4 h from cell seeding. The assay was tested in two separate cellular systems, namely, HeLa and MCF-7 cells. When using the one-well protocol with Akt phosphorylation as a model, the response to a number of agonists was the same as the response obtained using cells treated in a separate microplate, using a conventional lysate transfer approach. The assay procedure was automated, and quantitative pharmacological data on three known inhibitors of the PI3-kinase signaling pathway was obtained within 4 h from seeding cells, with six dispense steps, and a single wash cycle. Thus, the protocol affords a reliable means of assaying for cellular signaling events in different cell types, and is amenable to automation.

Christmas cheer or drear?

I once heard of a man who was given a brand new car for Christmas.

Decriminalising drugs.

RCN Congress last year saw one of its most volatile debates - around a matter for discussion on the issue of decriminalisation of drugs. There is no doubt this is an issue which can polarise opinion.