/ An extensive review of the published literature identified more than 150 case studies in which some aspect of resilience in freshwater systems was reported. Approximately 79% of systems studied were Iotic and the remainder lentic. Most of the stressor types were chemical with DDT (N = 29) and rotenone (N = 15) the most common. The most common nonchemical stressors were logging activity (N = 16), flooding (N = 8), dredging (N = 3), and drought (N = 7).The variety of endpoints to which recovery could be measured ranged from sparse data for phytoplankton (N = 13), periphyton (N = 6), and macrophytes (N = 8) to relatively more data for fish (N = 412) and macroinvertebrates (N = 698). Unfortunately the same characteristics were rarely measured consistently among sites. For example, with respect to fish, more than 30 different species were studied and recovery was measured in many ways, most commonly on the basis of: (1) first reappearance of the species, (2) return time of predisturbance densities, and (3) return time of predisturbance average individual size. Based on these criteria, all systems in these studies seem to be resilient to most disturbances with most recovery times being less than three years. Exceptions included when (1) the disturbance resulted in physical alteration of the existing habitat, (2) residual pollutants remained in the system, or (3) the system was isolated and recolonization was suppressed.The inherent capacity of an aquatic system to recover naturally from a stressor has been a neglected consideration in environmental assessments (Cairns 1978(Cairns , 1980. Primary reasons for this neglect are a lack of appreciation for the importance of recovery of an aquatic system following exposure to a stressor, and the lack of predisturbance data to properly assess whether a system has recovered. Most ecological research has focused on understanding how various stressors alter the chemical, physical, and biological function and structure of aquatic ecosystems. Once the effects are documented, funding generally has not been available to continue long-term studies of these systems (Likens 1983). Long-term studies of natural ecosystems are likely the only vehicle by which the recovery phase of a system can be understood.The lack of an appreciation for understanding the

A variety of G-protein-coupled receptor (GPCR) screening technologies have successfully partnered a number of GPCRs with their cognate ligands. GPCR-mediated β-arrestin recruitment is now recognized as a distinct intracellular signaling pathway, and ligand-receptor interactions may show a bias toward β-arrestin over classical GPCR signaling pathways. We hypothesized that the failure to identify native ligands for the remaining orphan GPCRs may be a consequence of biased β-arrestin signaling. To investigate this, we assembled 10 500 candidate ligands and screened 82 GPCRs using PathHunter β-arrestin recruitment technology. High-quality screening assays were validated by the inclusion of liganded receptors and the detection and confirmation of these established ligand-receptor pairings. We describe a candidate endogenous orphan GPCR ligand and a number of novel surrogate ligands. However, for the majority of orphan receptors studied, measurement of β-arrestin recruitment did not lead to the identification of cognate ligands from our screening sets. β-Arrestin recruitment represents a robust GPCR screening technology, and ligand-biased signaling is emerging as a therapeutically exploitable feature of GPCR biology. The identification of cognate ligands for the orphan GPCRs and the extent to which receptors may exist to preferentially signal through β-arrestin in response to their native ligand remain to be determined.

Chemotherapy that is used to treat human immunodeficiency virus type-1 (HIV-1) infection focuses primarily on targeting virally encoded proteins. However, the combination of a short retroviral life cycle and high mutation rate leads to the selection of drug-resistant HIV-1 variants. One way to address this problem is to inhibit non-essential host cell proteins that are required for viral replication. Here we show that the activity of HIV-1 integrase stimulates an ataxia-telangiectasia-mutated (ATM)-dependent DNA damage response, and that a deficiency of this ATM kinase sensitizes cells to retrovirus-induced cell death. Consistent with these observations, we demonstrate that a novel and specific small molecule inhibitor of ATM kinase activity, KU-55933, is capable of suppressing the replication of both wild-type and drug-resistant HIV-1.

Variation in pharmacology and function of ligands at species orthologs can be a confounding feature in understanding the biology and role of poorly characterized receptors. Substantial selectivity in potency of a number of GPR35 agonists has previously been demonstrated between human and rat orthologs of this G protein-coupled receptor. Via a bioluminescence resonance energy transfer-based assay of induced interactions between GPR35 and ␤-arrestin-2, addition of the mouse ortholog to such studies indicated that, as for the rat ortholog, murine GPR35 displayed very low potency for pamoate, whereas potency for the reference GPR35 agonist zaprinast was intermediate between the rat and human orthologs. This pattern was replicated in receptor internalization and G protein activation assays. The effectiveness and mode of action of two recently reported GPR35 antagonists, methyl-5-[(tert-butylcarbamothioylhydrazinylidene)methyl]-1-(2,4-difluorophenyl)pyrazole-4-carboxylate (CID-2745687) and 2-hydroxy-4-[4-(5Z)-5-[(E)-2-methyl-3-phenylprop-2-enylidene]-4-oxo-2-sulfanylidene-1,3-thiazolidin-3-yl]butanoylamino)benzoic acid (ML-145), were investigated. Both CID-2745687 and ML-145 competitively inhibited the effects at human GPR35 of cromolyn disodium and zaprinast, two agonists that share an overlapping binding site. By contrast, although ML-145 also competitively antagonized the effects of pamoate, CID-2745687 acted in a noncompetitive fashion. Neither ML-145 nor CID-2745687 was able to effectively antagonize the agonist effects of either zaprinast or cromolyn disodium at either rodent ortholog of GPR35. These studies demonstrate that marked species selectivity of ligands at GPR35 is not restricted to agonists and considerable care is required to select appropriate ligands to explore the function of GPR35 in nonhuman cells and tissues.