Single-molecule fingerprinting of protein-drug interaction using a funneled biological nanopore.

In drug discovery, efficient screening of protein-drug interactions (PDIs) is hampered by the limitations of current biophysical approaches. Here, we develop a biological nanopore sensor for single-molecule detection of proteins and PDIs using the pore-forming toxin YaxAB. Using this YaxAB nanopore, we demonstrate label-free, single-molecule detection of interactions between the anticancer Bcl-xL protein and small-molecule drugs as well as the Bak-BH3 peptide. The long funnel-shaped structure and nanofluidic characteristics of the YaxAB nanopore enable the electro-osmotic trapping of diverse folded proteins and high-resolution monitoring of PDIs. Distinctive nanopore event distributions observed in the two-dimensional (ΔI/Io-versus-IN) plot illustrate the ability of the YaxAB nanopore to discriminate individual small-molecule drugs bound to Bcl-xL from non-binders. Taken together, our results present the YaxAB nanopore as a robust platform for label-free, ultrasensitive, single-molecule detection of PDIs, opening up a possibility for low-cost, highly efficient drug discovery against diverse drug targets.

Could we prove the nursing outcomes utilising clinical data warehouse? Effectiveness of pressure ulcer intervention in Korean tertiary hospital.

The use of Clinical Data Warehouse (CDW)  for research and quality improvement has become more frequent in the last 10 years. In this study, we used CDW to determine the effectiveness of pressure ulcer interventions offered by ward nurses and wound care nursing specialists. A retrospective clinical outcomes study that utilise CDW has been carried out. We identified 1415 patients who were evaluated as pressure ulcer risk group from 1 July 2019 to 31 December 2019. Kaplan-Meier survival analyses were used to estimate the time to occurrence of pressure ulcers. We compared the survival curves of each group by applying the log-rank test for significance. The overall median time to occurrence for both groups was 13 days (95% CI range: 11-14 days). The control group showed a longer median time (14 days) to occurrence than the case group (12 days). In the pressure ulcer stage I, the case group showed a longer median time (14 days) to occurrence than the control group (8 days), indicating that the intervention provided by the wound care nursing specialist was effective in stage I, and delayed the occurrence of pressure ulcers. The findings may be used as preliminary data for the utilisation of the CDW in the field of nursing research in the future. Also, facilitating the accessibility of the wound care nursing specialist in the general wards should be effective to decrease the incidence rates.

MUL1-RING recruits the substrate, p53-TAD as a complex with UBE2D2-UB conjugate.

The RING domain of MUL1 (RINGMUL1 ) alone mediates ubiquitylation of the p53-transactivation domain (TADp53 ). To elucidate the mechanism underlying the simultaneous recruitment of UBE2D2 and the substrate TADp53 by RINGMUL1 , we determined the complex structure of RINGMUL1 :UBE2D2 and studied the interaction between RINGMUL1 and TADp53 in the presence of UBE2D2-UB thioester (UBE2D2~UB) mimetics. The RINGMUL1 -binding induced the closed conformation of UBE2D2S22R/C85S -UBK48R oxyester (UBE2D2RS -UBR OE ), and strongly accelerated its hydrolysis, which was suppressed by the additional N77A-mutation of UBE2D2. Interestingly, UBE2D2S22R/N77A/C85S -UBK48R oxyester (UBE2D2RAS -UBR OE ) already formed a closed conformation in the absence of RINGMUL1 . Although TADp53 exhibited weak binding for RINGMUL1 or UBE2D2 alone, its binding affinity was enhanced and even further for RINGMUL1 :UBE2D2 and RINGMUL1 :UBE2D2RAS -UBR OE , respectively. The recognition of TADp53 by RINGMUL1 as a complex with UBE2D2~UB is related to the multivalency of the binding events and underlies the ability of RINGMUL1 to ubiquitylate the intrinsically disordered protein, TADp53 .

Effect of silica coating on bond strength between a gold alloy and metal bracket bonded with chemically cured resin.

OBJECTIVE: The purpose of this study was to evaluate the effects of three different surface conditioning methods on the shear bond strength (SBS) of metal brackets bonded directly to gold alloy with chemically cured resin. METHODS: Two hundred ten type III gold alloy specimens were randomly divided into six groups according to the combination of three different surface conditioning methods (aluminum oxide sandblasting only, application of a metal primer after aluminum oxide sandblasting, silica coating and silanation) and thermocycling (with thermocycling, without thermocycling). After performing surface conditioning of specimens in accordance with each experimental condition, metal brackets were bonded to all specimens using a chemically cured resin. The SBS was measured at the moment of bracket debonding, and the resin remnants on the specimen surface were evaluated using the adhesive remnant index. RESULTS: Application of metal primer after aluminum oxide sandblasting yielded a higher bond strength than that with aluminum oxide sandblasting alone (p < 0.001), and silica coating and silanation yielded a higher bond strength than that with metal primer after aluminum oxide sandblasting (p < 0.001). There was no significant change in SBS after thermocycling in all groups. CONCLUSIONS: With silica coating and silanation, clinically satisfactory bond strength can be attained when metal brackets are directly bonded to gold alloys using a chemically cured resin.