Flossing technique for endovascular repair of a penetrating cerebrovascular injury: illustrative case.

BACKGROUND: A teenage boy who had been stabbed in the neck presented with an extracranial traumatic functional carotid artery occlusion that could not be crossed in an antegrade fashion. Endovascular repair depends on obtaining catheter access proximal and distal to an injury within the true lumen. OBSERVATIONS: The occlusion was treated with flossing technique via the posterior communicating artery. After successful recanalization from a retrograde approach, the carotid artery occlusion was treated with a covered stent. LESSONS: The flossing technique is well established in peripheral vascular disease and may be beneficial in certain cases in the neck vasculature when antegrade access is difficult to obtain. Recanalization of an occluded carotid artery from retrograde approach may be successful in cases of trauma from knife wounds.

Reverse micelle encapsulation as a model for intracellular crowding.

Reverse micelles are discrete nanoscale particles composed of a water core surrounded by surfactant. The amount of water within the core of reverse micelles can be easily manipulated to directly affect the size of the reverse micelle particle. The water loading capacity of reverse micelles varies with temperature, and water can be shed if reverse micelles are exposed to low temperatures. The use of water shedding from the reverse micelle provides precise and comprehensive control over the amount of water available to solvate host molecules. Proteins encapsulated within reverse micelles can be studied to determine the effects of confinement and excluded volume. The data presented here provide an important bridge between commonly employed dilute in vitro studies and studies of the effects of a crowded environment, as found in vivo. Ubiquitin was encapsulated within bis(2-ethylhexyl) sodium sulfosuccinate AOT reverse micelles under various degrees of confinement and was compared with an analogously reconstituted sample of ubiquitin in the commonly used molecular crowding agent bovine serum albumin. The effects of encapsulation were monitored using chemical shift perturbation analysis of the amide (1)H and (15)N resonances. The results also reconcile alternative interpretations of protein cold denaturation within reverse micelles.

Use of reverse micelles in membrane protein structural biology.

Membrane protein structural biology is a rapidly developing field with fundamental importance for elucidating key biological and biophysical processes including signal transduction, intercellular communication, and cellular transport. In addition to the intrinsic interest in this area of research, structural studies of membrane proteins have direct significance on the development of therapeutics that impact human health in diverse and important ways. In this article we demonstrate the potential of investigating the structure of membrane proteins using the reverse micelle forming surfactant dioctyl sulfosuccinate (AOT) in application to the prototypical model ion channel gramicidin A. Reverse micelles are surfactant based nanoparticles which have been employed to investigate fundamental physical properties of biomolecules. The results of this solution NMR based study indicate that the AOT reverse micelle system is capable of refolding and stabilizing relatively high concentrations of the native conformation of gramicidin A. Importantly, pulsed-field-gradient NMR diffusion and NOESY experiments reveal stable gramicidin A homodimer interactions that bridge reverse micelle particles. The spectroscopic benefit of reverse micelle-membrane protein solubilization is also explored, and significant enhancement over commonly used micelle based mimetic systems is demonstrated. These results establish the effectiveness of reverse micelle based studies of membrane proteins, and illustrate that membrane proteins solubilized by reverse micelles are compatible with high resolution solution NMR techniques.