Dislocation of ricin toxin A chains in human cells utilizes selective cellular factors.

Ricin is a potent A-B toxin that is transported from the cell surface to the cytosol, where it inactivates ribosomes, leading to cell death. Ricin enters cells via endocytosis, where only a minute number of ricin molecules reach the endoplasmic reticulum (ER) lumen. Subsequently, the ricin A chain traverses the ER bilayer by a process referred to as dislocation or retrograde translocation to gain access to the cytosol. To study the molecular processes of ricin A chain dislocation, we have established, for the first time, a human cell system in which enzymatically attenuated ricin toxin A chains (RTA(E177D) and RTA(Δ177-181)) are expressed in the cell and directed to the ER. Using this human cell-based system, we found that ricin A chains underwent a rapid dislocation event that was quite distinct from the dislocation of a canonical ER soluble misfolded protein, null Hong Kong variant of α(1)-antitrypsin. Remarkably, ricin A chain dislocation occurred via a membrane-integrated intermediate and utilized the ER protein SEL1L for transport across the ER bilayer to inhibit protein synthesis. The data support a model in which ricin A chain dislocation occurs via a novel strategy of utilizing the hydrophobic nature of the ER membrane and selective ER components to gain access to the cytosol.

Perioperative management of pheochromocytoma: focus on magnesium, clevidipine, and vasopressin.

The perioperative management of pheochromocytomas requires meticulous anesthetic care. There has been considerable progress in its management, recently 3 agents that may be particularly advantageous to the anesthetic team have been identified. Magnesium sulfate is readily available, cheap, safe, and effective for hemodynamic control before tumor resection. It has demonstrated efficacy in adults, children, and in rarer scenarios, such as pheochromocytoma resection in pregnancy and in pheochromocytoma crises. Although only recently entering clinical practice, clevidipine exhibits a pharmacologic profile of great interest, showing efficacy in the management of hypertensive crisis and providing rapid titration and precise hemodynamic control. Its application in the perioperative management of pheochromocytoma before tumor resection recently has been described and likely will expand in the near future. Vasopressin has demonstrated utility in the management of catecholamine-resistant shock after tumor resection. A familiarity with these 3 agents offers anesthesia providers further effective pharmacologic options for managing the hemodynamic challenges inherent to this population before and after tumor resection.

A Room-Temperature Ferroelectric Resonant Tunneling Diode.

Resonant tunneling is a quantum-mechanical effect in which electron transport is controlled by the discrete energy levels within a quantum-well (QW) structure. A ferroelectric resonant tunneling diode (RTD) exploits the switchable electric polarization state of the QW barrier to tune the device resistance. Here, the discovery of robust room-temperature ferroelectric-modulated resonant tunneling and negative differential resistance (NDR) behaviors in all-perovskite-oxide BaTiO3 /SrRuO3 /BaTiO3 QW structures is reported. The resonant current amplitude and voltage are tunable by the switchable polarization of the BaTiO3 ferroelectric with the NDR ratio modulated by ≈3 orders of magnitude and an OFF/ON resistance ratio exceeding a factor of 2 × 104 . The observed NDR effect is explained an energy bandgap between Ru-t2g and Ru-eg orbitals driven by electron-electron correlations, as follows from density functional theory calculations. This study paves the way for ferroelectric-based quantum-tunneling devices in future oxide electronics.

Ricin. Mechanisms of cytotoxicity.

Ricin is a heterodimeric protein produced in the seeds of the castor oil plant (Ricinus communis). It is exquisitely potent to mammalian cells, being able to fatally disrupt protein synthesis by attacking the Achilles heel of the ribosome. For this enzyme to reach its substrate, it must not only negotiate the endomembrane system but it must also cross an internal membrane and avoid complete degradation without compromising its activity in any way. Cell entry by ricin involves a series of steps: (i) binding, via the ricin B chain (RTB), to a range of cell surface glycolipids or glycoproteins having beta-1,4-linked galactose residues; (ii) uptake into the cell by endocytosis; (iii) entry of the toxin into early endosomes; (iv) transfer, by vesicular transport, of ricin from early endosomes to the trans-Golgi network; (v) retrograde vesicular transport through the Golgi complex to reach the endoplasmic reticulum; (vi) reduction of the disulphide bond connecting the ricin A chain (RTA) and the RTB; (vii) partial unfolding of the RTA to render it translocationally-competent to cross the endoplasmic reticulum (ER) membrane via the Sec61p translocon in a manner similar to that followed by misfolded ER proteins that, once recognised, are targeted to the ER-associated protein degradation (ERAD) machinery; (viii) avoiding, at least in part, ubiquitination that would lead to rapid degradation by cytosolic proteasomes immediately after membrane translocation when it is still partially unfolded; (ix) refolding into its protease-resistant, biologically active conformation; and (x) interaction with the ribosome to catalyse the depurination reaction. It is clear that ricin can take advantage of many target cell molecules, pathways and processes. It has been reported that a single molecule of ricin reaching the cytosol can kill that cell as a consequence of protein synthesis inhibition. The ready availability of ricin, coupled to its extreme potency when administered intravenously or if inhaled, has identified this protein toxin as a potential biological warfare agent. Therapeutically, its cytotoxicity has encouraged the use of ricin in 'magic bullets' to specifically target and destroy cancer cells, and the unusual intracellular trafficking properties of ricin potentially permit its development as a vaccine vector. Combining our understanding of the ricin structure with ways to cripple its unwanted properties (its enzymatic activity and promotion of vascular leak whilst retaining protein stability and important immunodominant epitopes), will also be crucial in the development of a long awaited protective vaccine against this toxin.

Spinning out a star.

Spinouts rarely take off; most, in fact, fall into one or more of four traps that doom them from the start. Some companies spin out ventures that are too close to the core of their businesses, in effect selling off their crown jewels. Sometimes, a parent company uses the spinout primarily to pawn off debt or expenses or to quickly raise external capital for itself. Other times, a company may try to spin out an area of its business that lacks one or more of the critical legs of a successful company--a coherent business model, say, or a solid financial base. And in many cases, parent companies can't bring themselves to sever their ownership ties and give up control of their spinouts. R.J. Reynolds, the tobacco giant, managed to avoid these traps when it successfully spun out a most unlikely venture, the pharmaceutical company Targacept. As the story illustrates, the problem with spinouts is similar to the problem of rich children. Their parents have the wherewithal to spoil them or shelter them or cling to them, but what they need is tough love and discipline--much the same discipline that characterizes successful start-ups. R.J. Reynolds recognized that it didn't know that much about the pharmaceutical business and couldn't merely try to spin out a small clone of itself. It had to treat the venture as if it were essentially starting from scratch, with a passionate entrepreneurial leader, a solid business plan, help from outside partners in the industry, and ultimately substantial venture backing. That these lessons are less obvious to executives contemplating spinning out ventures closer to their core businesses may be why so many spinouts fail.

Multiple Rib Stress Fractures.

In brief Stress fracture of the ribs, an uncommon overuse injury, may mimic recalcitrant intercostal or back muscle strain. This report describes a 36-year-old man who sustained multiple rib stress fractures after he took up golf. Fatigue of the serratus anterior muscle appeared to be the mechanism for his injury. Relative rest followed by strengthening exercises helped the patient return to pain-free activity.