Reassessment of the pharmacology of Sphingosine-1-phosphate S1P3 receptor ligands using the DiscoveRx PathHunter™ and Ca2+ release functional assays.

BACKGROUND AND PURPOSE: DiscoverRx's PathHunter™ assay measures GPCR agonist potency, via the recruitment of ß-arrestin, independent of the subtype of G(α) protein activated. This assay is frequently used in drug discovery although little is known about the agonist pharmacology generated. Here we have compared agonist potency, efficacy and affinity values obtained in PathHunter™ assays with those from more established radioligand binding and functional techniques. EXPERIMENTAL APPROACH: Using cells expressing the human sphingosine-1-phosphate S1P(3) receptor at four different densities, we compared pharmacological affinity and efficacy values of four structurally distinct ligands - FTY720-P, VPC24191, CYM5442 and the endogenous agonist S1P - obtained from competition binding, functional Ca(2+) release and PathHunter™ assays. KEY RESULTS: The pK(i) values for S1P were significantly different (9.34 ± 0.10 and 8.92 ± 0.15) in clones expressing different receptor levels using the binding assay. In the PathHunter™ and Ca(2+) assays, S1P and CYM5442 were full agonists, FTY720-P was a partial agonist, while the efficacy of VPC24191 could not be detected in PathHunter™ assays. VPC23019, previously described as a S1P(1/3) receptor antagonist, behaved as an S1P(3) receptor partial agonist in the Ca(2+) release assay. CONCLUSIONS AND IMPLICATIONS: Comparison of data from the PathHunter™ assay with binding and functional Ca(2+) assays suggest that PathHunter™ assays measured a different agonist-bound receptor conformation. While this assay has great utility in drug discovery, care must be taken as high-efficacy, low-affinity agonist compounds would not be detected. Therefore highly amplified, more traditional assays are necessary to identify agonists with low efficacy.

Working and scrounging by zebra finches in an operant task.

Groups of three zebra finches were housed in cages which could be adjoined to an operant chamber. Food was restricted and its availability was signalled by a red light projected behind an operant key (an autoshaping procedure). In addition to the shaping contingency, a single peck to the illuminated key, produced food immediately. Following the first peck to the illuminated key, by any subject in the group, the entire group was placed on a schedule of continuous reinforcement (CRF) wherein hopper operation was contingent upon a keypeck during the light. Comparison of individual's behavior revealed a differential pattern of operant working; one group member worked consistently, one worked periodically and one made few (if any) operant responses. All members consumed some reinforcers. The results suggest that zebra finches, under certain conditions, will form specialized labor roles. These data support similar demonstrations with primates (Chalmeau, R. and Galleau A., 1993. Behav. Processes, 28: 173-180), pigeons (Giraldeau, L. and Lefebvre L., 1987. Anim. Behav., 35: 387-394.) and rats (Thullier, F., Desor, D., Mos, J. and Krafft, B., 1992. Psychol. Behav., 53, 17-20).

Examining the relationship between Machiavellianism and paranoia.

To investigate the relationship between Machiavellianism and paranoid tendencies we administered the Mach IV scale and Fenigstein and Vanable's Paranoia scale to 150 students enrolled in 4 sections of a course, Introduction to Psychology, at a university in the Middle South. Scores on the 2 were significantly and positively correlated (Pearson r = .50). Men had a significantly higher mean score on Machiavellianism than women. Although men obtained a higher mean on the Paranoia scale than women, the difference was not statistically significant. Implications are discussed.

Adaptive communication systems for patients with mobility disorders.

An adaptive communication system has been developed for individuals with mobility disorders. It uses specialized computer software and hardware that compensate for this disability. For an individual with a motor-control impairment who is not able to use a keyboard effectively, a computer voice-recognition technology now removes this communication barrier. Speech-recognition systems consist of three basic components: speech processing, speech recognition, and speech understanding. The new Dragon Dictate (Dragon Systems, Inc., Newton, Mass.) is the first large-vocabulary speech recognition system in the personal computer industry that interactively learns a user's vocabulary and mode of speaking and responds to natural language rather than to limited sets of words. This speech-recognition system requires a microprocessor, a display monitor, a printer, and specific software packages, including word-processing and enhanced memory-management software. Important considerations in the use of this speech-recognition system include microphone positioning and training of the system. With the advent of this new voice-recognition computer system, another communication barrier between the disabled and society has been overcome.

Surgical needle holder damage to sutures.

Needle holder jaws with teeth produce distinct structural changes in synthetic sutures that cause a marked reduction in the suture breaking strength. This damage to the suture may account for disruptions of vascular and fascial wound closures in which continuous closure techniques are employed.

Large disposable trephines for circular excision biopsy of the skin.

Large disposable circular trephines have been developed specially for circular excision biopsy of the skin. This technique is a simple and practical alternative to the traditional method of excision. It has the advantages of aligning the incision in the direction of the maximum skin tensions and reducing the length of the final wound closure.