Small proteins or protein domains generally require disulfide bridges or metal sites for their stabilization. Here it is shown that the beta beta alpha architecture of zinc fingers can be reproduced in a 23-residue polypeptide in the absence of metal ions. The sequence was obtained through an iterative design process. A key feature of the final design is the incorporation of a type II' beta turn to aid in beta-hairpin formation. Nuclear magnetic resonance analysis reveals that the alpha helix and beta hairpin are held together by a defined hydrophobic core. The availability of this structural template has implications for the development of functional polypeptides.

Objective-Leukotriene B 4 (LTB 4 ), a product of the 5-lipoxygenase (5-LO) pathway of arachidonic acid metabolism, has been implicated in atherosclerosis. However, the molecular mechanisms for the atherogenic effect of LTB 4 are not well understood. This study is to determine candidate mechanisms. Method and Results-Primary human monocytes were treated with LTB 4 and the supernatant was analyzed for cytokine/chemokine production by an immuno-protein array. This analysis revealed a strong increase of the monocyte chemoattractant protein-1 (MCP-1), a proinflammatory cytokine. See page 1748Recent studies suggest a strong link between LTB 4 pathways with atherosclerosis. For example, human atherogenic plaques produce LTB 4 , 7 and expression of both BLT 1 and BLT 2 increases with the progression of atherosclerotic lesions. 8 Treatment of atherosclerosis-prone mice (apolipoprotein E [ApoE] or low-density lipoprotein receptor [LDLR]-deficient mice) with the BLT 1 -specific antagonist CP-105,696 9,10 markedly decreased lesion size. 11 Interestingly, the anti-atherogenic effects of CP-105,696 were diminished in mice deficient in the chemoattractant monocyte chemotactic protein-1 (MCP-1), 11 indicating a critical role for MCP-1 in mediating the LTB 4 atherogenic signals.MCP-1 is a prototype of the C-C chemokine ␤ subfamily and exhibits the most potent chemotactic activity for monocytes. 12 Overexpression of MCP-1 contributes to the development of atherosclerosis in mouse models. 13 Deficiency of either MCP-1 or its cognate high-affinity receptor C-C chemokine receptor 2 (CCR2) results in a marked decrease in atheromas and fewer monocytes in vascular lesions. 14,15 Additionally, therapeutic gene transfer of a dominantnegative MCP-1 mutant attenuated the development of early atherosclerosis and also limited progression of preexisting atherosclerotic lesions in ApoE-null mice. 16 Despite the critical role played by LTB 4 in atherogenesis, the molecular mechanisms for these activities are poorly understood. In this study, we investigated specifically whether LTB 4 regulates MCP-1 production in primary human monocytes to broaden the mechanistic understanding for LTB 4 -induced atherosclerosis. Our study shows that LTB 4 induced MCP-1 protein by several hundred-fold in primary human monocytes. LTB 4 induced MCP-1 mRNA by 500-fold Original

The discovery of vibegron, a potent and selective human β3-AR agonist for the treatment of overactive bladder (OAB), is described. An early-generation clinical β3-AR agonist MK-0634 (3) exhibited efficacy in humans for the treatment of OAB, but development was discontinued due to unacceptable structure-based toxicity in preclinical species. Optimization of a series of second-generation pyrrolidine-derived β3-AR agonists included reducing the risk for phospholipidosis, the risk of formation of disproportionate human metabolites, and the risk of formation of high levels of circulating metabolites in preclinical species. These efforts resulted in the discovery of vibegron, which possesses improved druglike properties and an overall superior preclinical profile compared to MK-0634. Structure-activity relationships leading to the discovery of vibegron and a summary of its preclinical profile are described.

We reviewed laboratory, clinical, and epidemiologic data on 35 patients from whom organisms belonging to the genus Roseomonas, a pink-pigmented gram-negative coccobacillus, were isolated over a 22-year period (1972-1994). Roseomonas strains were most commonly isolated from middle-aged women with one of several underlying conditions, including cancer and diabetes. Roseomonas was most commonly isolated from the blood, in association with clinical signs of sepsis. Approximately 60% of all isolates were judged to be of possible clinical significance, either as primary or secondary pathogens; 75% of all strains were recovered in pure culture. Roseomonas gilardii was the most frequently isolated species and was significantly associated with septicemia and underlying immunocompromised conditions; the species of 29% of all Roseomonas isolates could not be unequivocally identified with presently available differential tests. Genomospecies 5, currently an unnamed taxon within the genus Roseomonas, was primarily recovered as a commensal from young adults attending a sexually transmitted diseases clinic. The findings suggest that although this genus appears to have an overall low pathogenic potential for humans, Roseomonas species-in particular, R. gilardii-may be significant pathogens in persons with underlying medical complications.