Functional assays assay robustness

A PDE10A inhibitor may also have the potential to treat the cognitive symptoms of schizophrenia. The principal evidence for this claim is papaverine reversal of a PCP-induced deficit in the EDID-set shifting assay in rats [35]. This assay translates into human behavior in the form of the Wisconsin Card Sorting Test (WCST). EDID-set shifting is a test of executive function, a measure in which schizophrenics have a robust deficit. It has also been shown recently that papaverine is efficacious in the Novel Object Recognition cognition assay [36]. 

Protein microarrays have many potential applications in high-throughput analysis of protein function. However, simple, reproducible, and robust methods for array fabrication are required. Here we discuss the background to different routes to array fabrication and describe in detail one approach in which the purification and immobilization procedures are combined into a single step, dramatically simplifying the array fabrication process. We illustrate this approach by reference to the creation of an array of p53 variants, and discuss methods for assay and data analysis on such arrays. 

Efforts to test hypothesis two by biochemical reconstitution of a functional desaturase complex in vitro proved technically difficult and failed to provide a reproducible assay for functional expression of cloned lepidopteran desaturase sequences. In contrast, an in vivo expression system consisting of the yeast olel mutant and YEpOLEX plasmid confirmed hypothesis two and provided a technically facile and robust assay for determining the functional identities of many moth desaturase-encoding cDNAs. Particularly desirable features of this... 

CF-subjects with mild or severe lung disease and CBAVD subjects with one or two CFTR mutations (Fig. 6). To measure mutant CFTR function in vivo, nasal potential difference (NPD) recording techniques are used. This in vivo electrophysiology assay measures ion transport across the nasal membrane of human subjects. CFTR activity is isolated by addition of Cl" free media and /J-adrcncrgic agonists to increase cAMP signaling. The non-CF subjects respond with a robust increase in wild-type CFTR activity. In contrast, the response in subjects with no, mild and severe lung disease was 68 9% (44-88%), 9 6% (0-33%), and <1% of that observed for wild-type CFTR, respectively (Fig. 6 Table 4). 

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