"Plate cherry picking": a novel semi-sequential screening paradigm for cheaper, faster, information-rich compound selection

This work describes a novel semi-sequential technique for in silico enhancement of high-throughput screening (HTS) experiments now employed at Novartis. It is used in situations in which the size of the screen is limited by the readout (e.g., high-content screens) or the amount of reagents or tools (proteins or cells) available. By performing computational chemical diversity selection on a per plate basis (instead of a per compound basis), 25% of the 1,000,000-compound screening was optimized for general initial HTS. Statistical models are then generated from target-specific primary results (percentage inhibition data) to drive the cherry picking and testing from the entire collection. Using retrospective analysis of 11 HTS campaigns, the authors show that this method would have captured on average two thirds of the active compounds (IC(50) < 10 microM) and three fourths of the active Murcko scaffolds while decreasing screening expenditure by nearly 75%. This result is true for a wide variety of targets, including G-protein-coupled receptors, chemokine receptors, kinases, metalloproteinases, pathway screens, and protein-protein interactions. Unlike time-consuming "classic" sequential approaches that require multiple iterations of cherry picking, testing, and building statistical models, here individual compounds are cherry picked just once, based directly on primary screening data. Strikingly, the authors demonstrate that models built from primary data are as robust as models built from IC(50) data. This is true for all HTS campaigns analyzed, which represent a wide variety of target classes and assay types.     

In vitro rearing of Eucelatoria bryani: improvements and evaluation of factors affecting efficiency

In vitro rearing of Eucelatoria bryani Sabrosky (Diptera: Tachinidae) was made more efficient and economical. Absorbent cotton, used as a support to replace more expensive agar in an artificial medium, produced yields of adults equal to those reared on the agar-based medium. The weights of pupae from the cotton-supported medium were about 16% lighter than were the weights of pupae from the agar-based medium. Evaluation of diet volumes (100, 200 and 250 l) for individually reared flies revealed that highest adult yields (46.3%) were obtained when 200 l per maggot of agar-based diet were used in each well of microtiter plates; this is equivalent to 2,315 adults per liter of diet. The tachinids were small and the sizes were equal to those obtained when 10–15 maggots parasitize a single host. Costs could not be reduced by deleting free amino acids from the medium fed to older maggots because free amino acids are essential dietary ingredients for third instars of E. bryani.     

Prerequisites for oral immunization of free-ranging raccoons (Procyon lotor) with a recombinant rabies virus vaccine: study site ecology and bait system development.

A model baiting system suitable for the delivery of an oral rabies vaccine to free-ranging raccoons (Procyon lotor) was developed and tested on barrier islands in South Carolina (USA). Features of barrier island physiography and ecology were studied relative to selective bait deployment and site biosecurity. Capture-mark-recapture data were obtained from 228 raccoons. Raccoon density estimates, using a modified census assessment technique, were one raccoon per 1.8 to 2.7 ha. Mean (+/- SE) and range home area estimates of radio-collared raccoons were 84 (+/- 15.6) ha (27 to 176 ha) by a minimum convex polygon method and 138 (+/- 22.8) ha (43 to 241 ha), by a harmonic mean transformation method. Habitat utilization determinations of radio-collared raccoons were conducted to identify study areas to potentially maximize selectivity of bait towards raccoons and to reduce the absolute number of baits deployed. Island raccoons showed a habitat preference for maritime forest, maritime shrub and marsh areas. Additionally, there was no evidence of inter-island or mainland exchange of ear-tagged or radio-collared raccoons. A disease and mortality survey was conducted to identify baseline pathology and incidental lesions in the target raccoon population, prior to actual vaccination initiation. Thirty-eight percent of 30 clinically suspect raccoons sampled had intracytoplasmic eosinophilic inclusions diagnostic of canine distemper; no other lesions suggestive of viral etiologies were found. Serological surveys for raccoon poxvirus and rabies virus antibodies were negative. Antibody titers to canine adenovirus 1 and 2 indicated a moderate level of exposure (approximately 10 to 16%) in the raccoon population. Overall, 93 to 100% of placebo baits were consistently disturbed by 7 days post-bait deployment, and bait acceptance rates by raccoons ranged from 49 to 85%, by using a modular systems approach to select the optimum combination of bait attractant, biomarker, matrix, density, and distribution. These results suggest that a large proportion (up to 85%) of a free-ranging island raccoon population can be selectively and safely targeted, marked and monitored utilizing a proposed oral bait delivery system for recombinant or other rabies vaccines.     

A field trial to determine the feasibility of delivering oral vaccines to wild swine

A field study was conducted on Ossabaw Island, Georgia (USA) to determine the feasibility of delivering oral vaccines to wild swine (Sus scrofa). Baits were made of polymerbound fish meal and contained a gelatin capsule as a potential vaccine chamber. Two biomarkers, iophenoxic acid and tetracycline, were incorporated into each bait, and soured chicken mash was used as an attractant. Baits (n = 1,980) were distributed in a grid pattern on a 405-ha test site and monitored for animal disturbance. Within 72 hr, 88% of 393 monitored baits were gone, and observations of track-beds surrounding 100 baits indicated that at least 52% were taken by wild swine. Subsequent testing of 80 wild swine for the biomarkers revealed that 95% of the animals had consumed bait. Track-bed observations indicated that raccoons (Procyon lotor) were the only non-target animal that frequently took baits. Biomarker analyses indicated 44% of 16 raccoons tested had eaten bait. It was concluded that oral vaccine delivery to wild swine should be considered as a feasible method of control or eradication of pseudorabies and/or swine brucellosis in wild swine if effective vaccines become available.     

Lesions associated with pulmonary parasites in bobcats (Felis rufus) from Arkansas

Two of five bobcats (Felis rufus) from southwestern Arkansas had natural pulmonary infections of Paragonimus kellicotti and Filaroides rostratus. Pairs of P. kellicotti were found in spherical cyst-like structures approximately 1 cm in diameter. Filaroides rostratus were seen as serpentine pale white areas on the pleural surface and also firmly embedded in fibrous capsules in the pulmonary parenchyma. Histologic lesions associated with the presence of these parasites consisted primarily of a verminous bronchitis. Clinical signs or compromised pulmonary function were not associated with these infections.