Differential Scanning Fluorometry Signatures as Indicators of Enzyme Inhibitor Mode of Action: Case Study of Glutathione S-Transferase

Differential scanning fluorometry (DSF), also referred to as fluorescence thermal shift, is emerging as a convenient method to evaluate the stabilizing effect of small molecules on proteins of interest. However, its use in the mechanism of action studies has received far less attention. Herein, the ability of DSF to report on inhibitor mode of action was evaluated using glutathione S-transferase (GST) as a model enzyme that utilizes two distinct substrates and is known to be subject to a range of inhibition modes. Detailed investigation of the propensity of small molecule inhibitors to protect GST from thermal denaturation revealed that compounds with different inhibition modes displayed distinct thermal shift signatures when tested in the presence or absence of the enzyme's native co-substrate glutathione (GSH). Glutathione-competitive inhibitors produced dose-dependent thermal shift trendlines that converged at high compound concentrations. Inhibitors acting via the formation of glutathione conjugates induced a very pronounced stabilizing effect toward the protein only when GSH was present. Lastly, compounds known to act as noncompetitive inhibitors exhibited parallel concentration-dependent trends. Similar effects were observed with human GST isozymes A1-1 and M1-1. The results illustrate the potential of DSF as a tool to differentiate diverse classes of inhibitors based on simple analysis of co-substrate dependency of protein stabilization

Lawson criterion for ignition exceeded in an inertial fusion experiment

For more than half a century, researchers around the world have been engaged in attempts to achieve fusion ignition as a proof of principle of various fusion concepts. Following the Lawson criterion, an ignited plasma is one where the fusion heating power is high enough to overcome all the physical processes that cool the fusion plasma, creating a positive thermodynamic feedback loop with rapidly increasing temperature. In inertially confined fusion, ignition is a state where the fusion plasma can begin "burn propagation" into surrounding cold fuel, enabling the possibility of high energy gain. While "scientific breakeven" (i.e., unity target gain) has not yet been achieved (here target gain is 0.72, 1.37 MJ of fusion for 1.92 MJ of laser energy), this Letter reports the first controlled fusion experiment, using laser indirect drive, on the National Ignition Facility to produce capsule gain (here 5.8) and reach ignition by nine different formulations of the Lawson criterion

A neurosurgical view of anatomical evaluation of anterior C1–C2 for safer transoral odontoidectomy

An anatomical study for evaluation of anterior C1–C2. To provide essential anatomic data for safer transoral odontoidectomy. The surface dimensions of the atlas vertebra and the transoral approach for odontoidectomy have been described in detail. Anterior arcus of C1 must be drilled out to reach odontoid process for transoral odontoidectomy. The thickness of anterior ring of C1 has not been studied before. Sixty, dried adult atlas and 60 axis vertebrae and ten cadaveric craniocervical specimens were measured for the following: (1) bony drilling depth (BDD), the distance from the anterior wall of anterior ring of C1 to anterior wall of odontoid; (2) minimum drilling diameter (MDD), distance of minimum C1 anterior ring removal for odontoid resection on horizontal plane; (3) maximum bony drilling diameter (MBDD), distance of maximum C1 anterior ring removal for odontoid resection on horizontal plane. Lateral border of this diameter is limited by medial borders of the lateral mass; (4) the widest odontoid diameters (WOD) on coronal sections were measured. On 60 atlas and axis vertebrae, the BDD was 7.0 ± 1.2 mm on dry bones, the distance between the medial borders of the lateral mass (MBDD) was 16.1 ± 1.5 mm, and the WOD on coronal sections (WOD) was 9.8 ± 0.8 mm. On cadavers, the distance between the two edges of C1 anterior ring removal for odontoid resection (MDD) was 10.8 ± 1.1 mm and the WOD on coronal sections (WOD) was 10.1 ± 1.4 mm. An odontoid surgery through transoral approach is safe and feasible. A quantitative understanding of the anterior anatomy of C-1 and C-2 is necessary when considering transoral odontoid resection. In this study the authors define safe zones for anterior atlas and axis