SJS/TEN 2019: From science to translation.

Stevens-Johnson syndrome and toxic epidermal necrolysis (SJS/TEN) are potentially life-threatening, immune-mediated adverse reactions characterized by widespread erythema, epidermal necrosis, and detachment of skin and mucosa. Efforts to grow and develop functional international collaborations and a multidisciplinary interactive network focusing on SJS/TEN as an uncommon but high burden disease will be necessary to improve efforts in prevention, early diagnosis and improved acute and long-term management. SJS/TEN 2019: From Science to Translation was a 1.5-day scientific program held April 26-27, 2019, in Vancouver, Canada. The meeting successfully engaged clinicians, researchers, and patients and conducted many productive discussions on research and patient care needs.

Polymeric nanocarriers for siRNA delivery to murine macrophages.

This work investigates the interactions of a polycationic nanocarrier with siRNA and with cells in order to better understand the capabilities and limitations of the carrier. The polycationic nanocarriers are cross-linked copolymer nanoparticles synthesized in a single-step reaction using ARGET ATRP (activators regenerated by electron transfer atom transfer radical polymerization). The polycationic nanocarriers efficiently bind siRNA for polymer/siRNA mass ratios less than 1. A method to prepare fluorescently labeled polycationic nanocarriers is presented. The fluorescently labeled polycationic nanocarriers are used to investigate cellular internalization with RAW264.7 murine macrophage cells. Flow cytometry demonstrates that the uptake increased with nanoparticle concentration and incubation time. Confocal microscopy confirmed internalization of fluorescently labeled nanoparticles. The investigation of siRNA-induced knockdown demonstrates that higher concentrations of nanoparticles and siRNA are associated with increased knockdown. For the conditions tested in the knockdown experiments, the ARGET ATRP polycationic nanocarriers outperformed a commercially available Lipofectamine control.

Polycationic nanoparticles for siRNA delivery: comparing ARGET ATRP and UV-initiated formulations.

In this work, we develop and evaluate polycationic nanoparticles for the delivery of small interfering RNA (siRNA). Delivery remains a major challenge for translating siRNA to the clinic, and overcoming the delivery challenge requires effective siRNA delivery vehicles that meet the demands of the specific delivery strategy. Cross-linked polycationic nanoparticle formulations were synthesized using ARGET ATRP or UV-initiated polymerization. The one-step, one-pot, surfactant-stabilized monomer-in-water synthesis technique may provide a simpler and faster alternative to complicated, multistep techniques and an alternative to methods that rely on toxic organic solvents. The polymer nanoparticles were synthesized using the cationic monomer 2-(diethylamino)ethyl methacrylate, the hydrophobic monomer tert-butyl methacrylate to tune pH responsiveness, the hydrophilic monomer poly(ethylene glycol) methyl ether methacrylate to improve biocompatibility, and cross-linking agent tetraethylene glycol dimethacrylate to enhance colloidal stability. Four formulations were evaluated for their suitability as siRNA delivery vehicles in vitro with the human embryonic kidney cell line HEK293T or the murine macrophage cell line RAW264.7. The polycationic nanoparticles demonstrated efficient and rapid loading of the anionic siRNA following complexation. Confocal microscopy as well as flow cytometry analysis of cells treated with polycationic nanoparticles loaded with fluorescently labeled siRNA demonstrated that the polycationic nanoparticles promoted cellular uptake of fluorescently labeled siRNA. Knockdown experiments using polycationic nanoparticles to deliver siRNA demonstrated evidence of knockdown, thus demonstrating potential as an alternative route to creating polycationic nanoparticles.

Oral delivery of small RNA and DNA.

The promise of short RNA or DNA segments such as siRNAs, antisense oligonucleotides, and transcription factor decoys to treat disease has prompted nearly 40 clinical trials for RNA interference (RNAi)-based therapeutics and more than 100 clinical trials for antisense oligonucleotide-based technologies; in fact, there are promising in vivo and preclinical studies for many hundreds of technologies. Current treatment strategies are largely injection-based, so effective oral delivery platforms for oligonucleotides may result in improved patient comfort and compliance. We analyze recently developed oral delivery methods for short RNA and DNA segments.

Fibromyalgia: revisiting the literature.

Fibromyalgia has a distinct clinical presentation. With no distinct characteristic beyond the presence of 11 or more tender points and chronic pain in all four quadrants of the body, it represents one extreme of a normal distribution of pain states. Research exploration utilizing the ACR criteria has not found solid empirical evidence to link the finding of multiple tender points to a specific pathological process. These points may be present as a concomitant finding with psychological disease states but they have not lead to further etiological understanding. Measurement of outcomes is difficult and the prognosis for patients in the specialty care setting is poor, however in the general population the prognosis is variable, and includes improvement without treatment. The success of treatments has been limited mainly to helping patients improve their ability to cope with, but not to eliminate the tender points.