Synthesis of trifold-labeled versatile reagent [3,5-13 C2 ,4-15 N]4-phenyl-1,2,4-triazoline-3,5-dione.

Triazolinediones are an important class of derivatization agents that have found application in various research disciplines. Their unique reactivity often allows precise and selective tagging of relevant molecular scaffolds to facilitate structural elucidation, tracking in biological systems, and stabilization of labile compounds. Recent research efforts mainly focused on the development of novel fluorescent and ionizable or isotopically labeled tags improving the quantification and identification of the parent molecule by suitable analytical methods. However, these concepts often lack the ability to improve properties facilitating the analysis by nuclear magnetic resonance (NMR) spectroscopy. We herein describe the first synthesis of 13 C and 15 N labeled [3,5-13 C2 ,4-15  N]4-phenyl-1,2,4-triazoline-3,5-dione utilizing the Cookson/Zinner-Deucker synthesis of urazoles. The introduced isotopic labels are ideally suited to support the structural elucidation of unknown and complex derivatization mixtures by NMR, thereby exploiting the increased sensitivity of detecting long-range JHC and additional JCC and JCN couplings within the derivatized compounds of interest.

Femtosecond Plasmonic Laser Nanosurgery (fs-PLN) mediated by molecularly targeted gold nanospheres at ultra-low pulse fluences.

Plasmonic Laser Nanosurgery (PLN) is a novel photomodification technique that exploits the near-field enhancement of femtosecond (fs) laser pulses in the vicinity of gold nanoparticles. While prior studies have shown the advantages of fs-PLN to modify cells, further reduction in the pulse fluence needed to initiate photomodification is crucial to facilitate deep-tissue treatments. This work presents an in-depth study of fs-PLN at ultra-low pulse fluences using 47 nm gold nanoparticles, conjugated to antibodies that target the epithelial growth factor receptor and excited off-resonance using 760 nm, 270 fs laser pulses at 80 MHz repetition rate. We find that fs-PLN can optoporate cellular membranes with pulse fluences as low as 1.3 mJ/cm2, up to two orders of magnitude lower than those used at lower repetition rates. Our results, corroborated by simulations of free-electron generation by particle photoemission and photoionization of the surrounding water, shed light on the off-resonance fs-PLN mechanism. We suggest that photo-chemical pathways likely drive cellular optoporation and cell damage at these off-resonance, low fluence, and high repetition rate fs-laser pulses, with clusters acting as local concentrators of ROS generation. We believe that the low fluence and highly localized ROS-mediated fs-PLN approach will enable targeted therapeutics and cancer treatment.

Catalyst Repurposing Sequential Catalysis by Harnessing Regenerated Prolinamide Organocatalysts as Transfer Hydrogenation Ligands.

A catalyst repurposing strategy based on a sequential aldol addition and transfer hydrogenation giving access to enantiomerically enriched α-hydroxy-γ-butyrolactones is described. The combination of a stereoselective, organocatalytic step, followed by an efficient catalytic aldehyde reduction induces an ensuing lactonization to provide enantioenriched butyrolactones from readily available starting materials. By capitalizing from the capacity of prolineamides to act as both an organocatalyst and a transfer hydrogenation ligand, catalyst repurposing allowed the development of an operationally simple, economic, and efficient sequential catalysis approach.

Symplectic homology product via Legendrian surgery.

This research announcement continues the study of the symplectic homology of Weinstein manifolds undertaken by the authors [Bourgeois F, Ekholm T, Eliashberg Y (2009) arXiv:0911.0026] where the symplectic homology, as a vector space, was expressed in terms of the Legendrian homology algebra of the attaching spheres of critical handles. Here, we express the product and Batalin-Vilkovisky operator of symplectic homology in that context.

Equivalent loads for two-dimensional distributed anisotropic piezoelectric transducers with arbitrary shapes attached to thin plate structures.

When a voltage is applied across the electrodes of a flat piezoelectric transducer attached to a thin plate structure, the transducer acts as equivalent loads applied to the host plate structure. In this paper, analytical expressions of these equivalent loads are derived for the general case of an orthotropic piezoelectric actuator using Hamilton's principle and two different mathematical approaches leading to the same results: Green's theorem and derivation using the theory of distributions. The equivalent loads are a function of the material properties as well as the normal to the contour of the transducer. Examples of applications to simple geometric shapes (triangle, rectangle, and circle) are given.

Ultrafast laser nanosurgery in microfluidics for genome-wide screenings.

The use of ultrafast laser pulses in surgery has allowed for unprecedented precision with minimal collateral damage to surrounding tissues. For these reasons, ultrafast laser nanosurgery, as an injury model, has gained tremendous momentum in experimental biology ranging from in vitro manipulations of subcellular structures to in vivo studies in whole living organisms. For example, femtosecond laser nanosurgery on such model organism as the nematode Caenorhabditis elegans has opened new opportunities for in vivo nerve regeneration studies. Meanwhile, the development of novel microfluidic devices has brought the control in experimental environment to the level required for precise nanosurgery in various animal models. Merging microfluidics and laser nanosurgery has recently improved the specificities and increased the speed of laser surgeries enabling fast genome-wide screenings that can more readily decode the genetic map of various biological processes.

Femtosecond laser nanoaxotomy properties and their effect on axonal recovery in C. elegans.

We present a study characterizing the properties of femtosecond laser nanosurgery applied to individual axons in live Caenorhabditis elegans (C. elegans) using nano-Joule laser pulses at 1 kHz repetition rate. Emphasis is placed on the characterization of the damage threshold, the extent of damage, and the statistical rates of axonal recovery as a function of laser parameters. The ablation threshold decreases with increasing number of pulses applied during nanoaxotomy. This dependency suggests the existence of an incubation effect. In terms of extent of damage, the energy per pulse is found to be a more critical parameter than the number of pulses. Axonal recovery improves when surgery is performed using a large number of low energy pulses.

Quantitative ultrasound method to detect and monitor laser-induced cavitation bubbles.

An ultrasound technique to measure the spatial and temporal behavior of the laser-induced cavitation bubble is introduced. The cavitation bubbles were formed in water and in gels using a nanosecond pulsed Nd:YAG laser operating at 532 nm. A focused, single-element, 25-MHz ultrasound transducer was employed both to detect the acoustic emission generated by plasma expansion and to acoustically probe the bubble at different stages of its evolution. The arrival time of the passive acoustic emission was used to estimate the location of the cavitation bubble's origin and the time of flight of the ultrasound pulse-echo signal was used to define its spatial extent. The results of ultrasound estimations of the bubble size were compared and found to be in agreement with both the direct optical measurements of the stationary bubble and the theoretical estimates of bubble dynamics derived from the well-known Rayleigh model of a cavity collapse. The results of this study indicate that the proposed quantitative ultrasound technique, capable of detecting and accurately measuring laser-induced cavitation bubbles in water and in a tissue-like medium, could be used in various biomedical and clinical applications.

Ultrasound measurements of cavitation bubble radius for femtosecond laser-induced breakdown in water.

A recently developed ultrasound technique is evaluated by measuring the behavior of a cavitation bubble that is induced in water by a femtosecond laser pulse. The passive acoustic emission during optical breakdown is used to estimate the location of the cavitation bubble's origin. In turn, the position of the bubble wall is defined based on the active ultrasonic pulse-echo signal. The results suggest that the developed ultrasound technique can be used for quantitative measurements of femtosecond laser-induced microbubbles.

Femtosecond laser nanoaxotomy lab-on-a-chip for in vivo nerve regeneration studies.

A thorough understanding of nerve regeneration in Caenorhabditis elegans requires performing femtosecond laser nanoaxotomy while minimally affecting the worm. We present a microfluidic device that fulfills such criteria and can easily be automated to enable high-throughput genetic and pharmacological screenings. Using the 'nanoaxotomy' chip, we discovered that axonal regeneration occurs much faster than previously described, and notably, the distal fragment of the severed axon regrows in the absence of anesthetics.

Femtosecond laser nanoaxotomy properties and their effect on axonal recovery in C. elegans.

We present a study characterizing the properties of femtosecond laser nanosurgery applied to individual axons in live Caenorhabditis elegans (C. elegans) using nano-Joule laser pulses at 1 kHz repetition rate. Emphasis is placed on the characterization of the damage threshold, the extent of damage, and the statistical rates of axonal recovery as a function of laser parameters. The ablation threshold decreases with increasing number of pulses applied during nanoaxotomy. This dependency suggests the existence of an incubation effect. In terms of extent of damage, the energy per pulse is found to be a more critical parameter than the number of pulses. Axonal recovery improves when surgery is performed using a large number of low energy pulses.

Information processing and movement optimization during development: kinematics of cyclical pointing in 5- to 11-year-old children.

The authors studied the development of movement control in speed-accuracy tradeoff conditions in children aged 5-11 years and in adults performing cyclical pointings. Twelve difficulty levels (IDs), ranging from 2 to 6.58 bits, were defined (P. M. Fitts, 1954). Peak and time to peak velocity, acceleration, and deceleration, and acceleration profiles as a function of hand position (Hooke's portraits) were analyzed. Movement time decreased with age and was less affected by ID. Peak velocity and acceleration increased, acceleration and deceleration were decreasingly time consuming, and movement profiles turned to increased harmonicity with age and task easiness. Nevertheless, the developmental trends differed between parameters. Gain in velocity seemed chiefly dependent on improved muscular cooperation patterns before 7 years of age and on improved information processing from age 7 onward; achievement of an optimized strategy in the speed-accuracy tradeoff occurred at age 11 years.