A Cyclic Mimic of HIV Tat Differentiates Similar TAR RNAs on the Basis of Distinct Dynamic Behaviors.

Efforts toward the development of RNA-based drug leads have been challenging because of the complexity and dynamic nature of RNA structures as therapeutic targets. The transactivation response (TAR) RNA and cognate Tat protein of HIV have long been recognized as promising antiviral targets, and recent works have identified potentially potent inhibitors of the viral RNA-protein interaction. A new class of such inhibitors, conformationally constrained cyclic peptide mimetics of Tat, has been demonstrated to inhibit the HIV life cycle. We have previously probed the complexity and dynamics of TAR RNAs in their free states, as well as conformational shifting by various peptide and small molecule ligands. In this work, we have used an ultrafast dynamics approach to probe the interactions between TAR RNAs and one of the representatives of cyclic peptide inhibitors, L22. Our studies demonstrated that cyclic L22 specifically recognizes TAR RNAs with a unique single binding site compared to two binding sites for linear Tat protein. Although both Tat and L22 bind to the TAR RNAs as a ß-hairpin structure, cyclization in L22 allows it to be a more efficient ligand from a population shifting perspective. This study provided unique insights into drug design with desired properties to differentiate similar structures based on distinct dynamic behaviors.

An efficient and general approach to beta-functionalized ketones.

[structure: see text]. The oxidation of selected anions (N3-, SCN-, I-, and Br-) by ceric ammonium nitrate (CAN) in the presence of substituted cyclopropyl alcohols provides a novel approach to beta-functionalized ketones. The protocol has a number of advantages including short reaction times, ease of reagent handling, and mild, neutral reaction conditions. Overall, this method provides an alternative pathway to important starting materials and intermediates in organic synthesis.

Detecting bacterial colonization of implanted orthopaedic devices by ultrasonication.

Glycocalyx-producing bacteria have been observed on orthopaedic devices that were removed for reasons other than infection. It has been suggested that the bacteria adhere to foreign surfaces within a biofilm and elude standard culture techniques. The authors adapted previously used ultrasonication protocols that disrupt the surface biofilm before culturing removed orthopaedic devices from patients without clinical evidence of infection. Patients having revision total joint arthroplasty of the hip or knee who lacked current or prior clinical evidence of infection were studied prospectively. During surgery, the femoral component and a corresponding control femoral implant were placed in separate sterile bags of saline. The implant and saline combination was placed in an ultrasonication bath for 30 minutes at 60 Hz. The saline solution was passed through a 0.45-microm pore filter, and the filter residue was cultured on sheep blood agar. None of the 21 implants yielded positive culture on routine microbiologic testing. However, using the ultrasonication protocol, a coagulase-negative Staphylococcus grew from one of the removed implants. Numerous total joint implant failures that are attributed to aseptic loosening may be a result of subclinical infection from bacteria within a biofilm. The current study supports the concept that biofilm-protected bacterial colonization of implants may occur without overt signs of infection and ultrasonication can be used to enhance identification of these bacteria.