Transcriptome-Wide, Unbiased Profiling of Ribonuclease Targeting Chimeras.

Various approaches have been developed to target RNA and modulate its function with modes of action including binding and cleavage. Herein, we explored how small molecule binding is correlated with cleavage induced by heterobifunctional ribonuclease targeting chimeras (RiboTACs), where RNase L is recruited to cleave the bound RNA target, in a transcriptome-wide, unbiased fashion. Only a fraction of bound targets was cleaved by RNase L, induced by RiboTAC binding. Global analysis suggested that (i) cleaved targets generally form a region of stable structure that encompasses the small molecule binding site; (ii) cleaved targets have preferred RNase L cleavage sites nearby small molecule binding sites; (iii) RiboTACs facilitate a cellular interaction between cleaved targets and RNase L; and (iv) the expression level of the target influences the extent of cleavage observed. In one example, we converted a binder of LGALS1 (galectin-1) mRNA into a RiboTAC. In MDA-MB-231 cells, the binder had no effect on galectin-1 protein levels, while the RiboTAC cleaved LGALS1 mRNA, reduced galectin-1 protein abundance, and affected galectin-1-associated oncogenic cellular phenotypes. Using LGALS1, we further assessed additional factors including the length of the linker that tethers the two components of the RiboTAC, cellular uptake, and the RNase L-recruiting module on RiboTAC potency. Collectively, these studies may facilitate triangulation of factors to enable the design of RiboTACs.

Sulfidated nanoscale zero-valent iron derived from iron sludge for tetracycline removal: Role of sulfur and iron in reactivity and mechanisms.

Iron sludge, produced during the drinking water treatment process, can be recycled as potential iron resource to create environmental functional material. In this study, sulfur-iron composites derived from iron sludge (S-Fe composites) was synthesized through sulfidation and carbonization, and used for the tetracycline (TC) removal under aerobic and anoxic conditions. The reactivities of these as-prepared products were strongly depended on pyrolysis temperatures. In particular, sulfidated nanoscale zero-valent iron loaded on carbon (S-nFe0@CIS) carbonized at 800 °C exhibited the highest TC removal efficiency with 86.6% within 30 min at circumneutral pH compared with other S-Fe composites. The crystalline structure of α-Fe0, FeSx and S0 as main active sites in S-nFe0@CIS promoted the degradation of TC. Moreover, the Fe/S molar ratios significantly affected the TC removal rates, which reached the best value as the optimal S/Fe of 0.27. The results illustrated that the optimized extent of sulfidation could facilitate electron transfer from nFe0 towards contaminants and accelerate Fe(III)/Fe(II) cycle in reaction system compared to bared nFe0@CIS. We revealed that removal of TC by S-nFe0@CIS in the presence of dissolved oxygen (DO) is mainly attributed to oxidation, adsorption and reduction pathways. Their contribution to TC removal were 31.6%, 25.2% and 28.8%, respectively. Furthermore, this adsorption-oxygenation with the formation of S-nFe0@CIS-TC* complexes was a surface-mediated process, in which DO was transformed by the structural FeSx on complex surface to •OH with the generation of H2O2 intermediate. The intermediates of TC and toxicity analysis indicate that less toxicity products generated through degradation process. This study provides a new reclamation of iron sludge and offers a new insight into the TC removal by S-nFe0@CIS under aerobic conditions.

Programming inactive RNA-binding small molecules into bioactive degraders.

Target occupancy is often insufficient to elicit biological activity, particularly for RNA, compounded by the longstanding challenges surrounding the molecular recognition of RNA structures by small molecules. Here we studied molecular recognition patterns between a natural-product-inspired small-molecule collection and three-dimensionally folded RNA structures. Mapping these interaction landscapes across the human transcriptome defined structure-activity relationships. Although RNA-binding compounds that bind to functional sites were expected to elicit a biological response, most identified interactions were predicted to be biologically inert as they bind elsewhere. We reasoned that, for such cases, an alternative strategy to modulate RNA biology is to cleave the target through a ribonuclease-targeting chimera, where an RNA-binding molecule is appended to a heterocycle that binds to and locally activates RNase L1. Overlay of the substrate specificity for RNase L with the binding landscape of small molecules revealed many favourable candidate binders that might be bioactive when converted into degraders. We provide a proof of concept, designing selective degraders for the precursor to the disease-associated microRNA-155 (pre-miR-155), JUN mRNA and MYC mRNA. Thus, small-molecule RNA-targeted degradation can be leveraged to convert strong, yet inactive, binding interactions into potent and specific modulators of RNA function.

The emission quenching of upconversion nanoparticles coated with amorphous silica by fluorescence resonance energy transfer: A mercury-sensing nanosensor excited by near-infrared radiation.

In this paper, a rhodamine derivative was synthesized as a probe for Hg(II) detection. Its spectral response and sensing mechanism towards Hg(II) were discussed carefully. It was found that its absorption and emission were increased by Hg(II), via a direct bonding stoichiometry of 1:1. Its association constant was determined with absorption titration as 2.59 × 105 M-1, which suggested a coordination procedure between Hg(II) and this rhodamine probe. It showed good selectivity towards Hg(II) over competing metal cations, no increased emission or absorption was observed in the presence of interfering metal cations. It was then covalently grafted onto silica (SiO2)-encapsulated upconversion nanoparticles (UCNPs). Upon near-infrared (NIR) excitation (980 nm), RHO accepted energy from these UCNPs through a FRET (fluorescence resonance energy transfer) procedure, quenching their upconversion emission. A sensing response towards Hg(II) was thus constructed. Good linearity and selectivity were still preserved in this composite sample. On the other hand, this work found a different phenomenon from literature cases, which was the emission absence of rhodamine emission in this composite structure. Detailed analysis suggested that rhodamine emission absence was caused by its self-quenching effect.

Functional Results after Intramuscular Ulnar Nerve Anterior Transposition for Young Adults Patients.

Background: This study investigated the functional outcomes of intramuscular ulnar nerve transposition (IMUNT) in young adults with cubital tunnel syndrome (CuTS). Methods: This retrospective study enrolled 37 military soldiers on active duty diagnosed with and treated for CuTS to determine the compression sites, complication rate, and postoperative results. Patient outcomes were analyzed using the Disability of the Arm, Shoulder, and Hand (DASH) questionnaire and the Bishop-Kleinman rating scales. Results: Patient outcomes were analyzed after a mean follow-up duration of 26.1 (22-29) months for 37 extremities. DASH scores improved from 38.7 (range, 13-63 points) preoperatively to 5.8 (range, 0-18 points) postoperatively. Patient improvement was statistically significant (p < 0.05). Based on the 12-point Bishop-Kleinman rating system, 30 (82.1%) patients were graded as excellent; five (13.5%) as good, and two (5.4%) as failed outcomes. Statistically significant improvements in both key pinch and grip strength were noted. Complications included one case of transient neuroparaxias of the medial antebrachial cutaneous nerve and one case of hematoma formation. Conclusions: We consider intramuscular ulnar nerve transposition to be a satisfactory procedure for CuTS. The procedure enhances upper limb function, thus allowing the patients to resume their physically demanding work with minimal complications.