The cytoplasmic domain of the AAA+ protease FtsH is tilted with respect to the membrane to facilitate substrate entry.

AAA+ proteases are degradation machines that use ATP hydrolysis to unfold protein substrates and translocate them through a central pore toward a degradation chamber. FtsH, a bacterial membrane-anchored AAA+ protease, plays a vital role in membrane protein quality control. How substrates reach the FtsH central pore is an open key question that is not resolved by the available atomic structures of cytoplasmic and periplasmic domains. In this work, we used both negative stain TEM and cryo-EM to determine 3D maps of the full-length Aquifex aeolicus FtsH protease. Unexpectedly, we observed that detergent solubilization induces the formation of fully active FtsH dodecamers, which consist of two FtsH hexamers in a single detergent micelle. The striking tilted conformation of the cytosolic domain in the FtsH dodecamer visualized by negative stain TEM suggests a lateral substrate entrance between the membrane and cytosolic domain. Such a substrate path was then resolved in the cryo-EM structure of the FtsH hexamer. By mapping the available structural information and structure predictions for the transmembrane helices to the amino acid sequence we identified a linker of ∼20 residues between the second transmembrane helix and the cytosolic domain. This unique polypeptide appears to be highly flexible and turned out to be essential for proper functioning of FtsH as its deletion fully eliminated the proteolytic activity of FtsH.

Contextual biases in the interpretation of auditory evidence.

Noisy recordings of dialogue often serve as evidence in criminal proceedings. The present article explores the ability of two types of contextual information, currently present in the legal system, to bias subjective interpretations of such evidence. The present experiments demonstrate that the general context of the legal system and the presence of transcripts of the recorded speech are both able to bias interpretations of degraded & benign recordings into interpretable & incriminating. Furthermore we demonstrate a curse of knowledge whereby people become miscalibrated to the true quality of degraded recordings when provided transcripts. Current methods of dealing with auditory evidence are insufficient to mollify the effects of biasing information within the criminal justice system.

Filling defects at CT colonography: pseudo- and diminutive lesions (the good), polyps (the bad), flat lesions, masses, and carcinomas (the ugly).

Numerous filling defects may be detected in the colon during interpretation of data sets obtained with computed tomographic (CT) colonography. A series of 230 patients were evaluated with thin-section multidetector row CT colonography immediately before conventional colonoscopy. In all cases, the interpreting radiologist and gastroenterologist reviewed the imaging findings as well as the results of histologic analysis of biopsy specimens to determine the causes of filling defects. In many cases, the cause of a filling defect can be confidently determined at CT colonography by using combinations of two- and three-dimensional images. However, lesions will occasionally be indeterminate because of overlapping features and will require further evaluation with endoscopy. With knowledge of the morphologic and attenuation characteristics of the various filling defects in the colon, one should be able to differentiate those filling defects detected at CT colonography that require no further evaluation from those that require endoscopic interrogation.