ITDetect: a method to detect internal tandem duplication of FMS-like tyrosine kinase (FLT3) from next-generation sequencing data with high sensitivity and clinical application.

Internal tandem duplication (ITD) of the FMS-like tyrosine kinase (FLT3) gene is associated with poor clinical outcomes in patients with acute myeloid leukemia. Although recent methods for detecting FLT3-ITD from next-generation sequencing (NGS) data have replaced traditional ITD detection approaches such as conventional PCR or fragment analysis, their use in the clinical field is still limited and requires further information. Here, we introduce ITDetect, an efficient FLT3-ITD detection approach that uses NGS data. Our proposed method allows for more precise detection and provides more detailed information than existing in silico methods. Further, it enables FLT3-ITD detection from exome sequencing or targeted panel sequencing data, thereby improving its clinical application. We validated the performance of ITDetect using NGS-based and experimental ITD detection methods and successfully demonstrated that ITDetect provides the highest concordance with the experimental methods. The program and data underlying this study are available in a public repository.

Subtrochanteric Insufficiency Fracture Occurring 5 Years after Surgery at the Steinmann Pin Insertion Site for Fracture Reduction.

Background and Objectives: Steinmann pins are commonly used in orthopedics, with a low rate of complications. However, thermal osteonecrosis may occur when a pin is inserted using a drill. There have been no reports on late-onset fractures at the Steinmann pin insertion site. Materials and Methods: A 32-year-old man who underwent surgery for a femoral shaft fracture 5 years ago complained of proximal thigh pain 1 month after the removal of the internal device. On physical examination, the patient showed a limping gait due to pain, and tenderness was observed on the lateral aspect of the proximal thigh. Magnetic resonance imaging was performed because the symptoms did not improve, despite conservative treatment. A new fracture line was observed in the lateral cortical bone of the proximal femur. It was found that a fracture occurred at the site where the Steinmann pin was inserted for a closed reduction at the time of the first operation. The patient was instructed to limit weight bearing and to use crutches while walking. Parathyroid hormone was additionally administered to promote bone formation. Results: Six months after diagnosis, a complete union was achieved at the subtrochanteric fracture site, and the patient's pain subsided. Conclusions: A fracture that occurs as a late onset at the provisional Steinmann pin insertion site is an extremely rare complication; however, orthopedic surgeons must consider this possibility and make more efforts to lower the occurrence of thermal damage. In addition, if the patient complains of pain in the region where the pin was inserted after surgery, surgeons should spare no effort to determine whether a new fracture has occurred.

Nanofluidic chip for liquid TEM cell fabricated by parylene and silicon nitride direct bonding.

Despite the importance of nanofluidic transmission electron microscope (TEM) chips, a simple fabrication method has yet to be developed due to the difficulty of wafer bonding techniques using a nanoscale thick bonding layer. We present a simple and robust wafer scale bonding technique using parylene as a bonding layer. A nanoscale thick parylene layer was deposited on a silicon nitride (SiN) wafer and patterned to construct nanofluidic channels. The patterned parylene layer was directly bonded to another SiN wafer by thermal surface activation and bonding, with a bonding strength of ∼3 MPa. Fourier transform infrared spectroscopy showed that carbon-oxygen bonds were generated by thermal activation. We demonstrated TEM imaging of gold nanoparticles suspended in liquid using the fabricated nanofluidic chip.

Direct transfer of multilayer graphene grown on a rough metal surface using PDMS adhesion engineering.

The direct transfer of graphene using polydimethylsiloxane (PDMS) stamping has advantages such as a 'pick-and-place' capability and no chemical residue problems. However, it is not easy to apply direct PDMS stamping to graphene grown via chemical vapor deposition on rough, grainy metal surfaces due to poor contact between the PDMS and graphene. In this study, graphene consisting of a mixture of monolayers and multiple layers grown on a rough Ni surface was directly transferred without the use of an adhesive layer. Liquid PDMS was cured on graphene to effect a conformal contact with the graphene. A fast release of graphene from substrate was achieved by carrying out wet-etching-assisted mechanical peeling. We also carried out a thermal post-curing of PDMS to control the level of adhesion between PDMS and graphene and hence facilitate a damage-free release of the graphene. Characterization of the transferred graphene by micro-Raman spectroscopy, SEM/EDS and optical microscopy showed neither cracks nor contamination from the transfer. This technique allows a fast and simple transfer of graphene, even for multilayer graphene grown on a rough surface.

Formvar Assisted Graphene Transfer for Graphene TEM Grid.

We report a formvar assisted transfer method to make graphene TEM grids having single layer graphene. Our method uses formvar instead of PMMA as the polymer support and produces clean graphene. Formvar was easily removed by dissolving in chloroform and did not leave residues after removal because of the lower the molecular weight. Our transfer method achieved the higher average coverage (about 55%) than direct transfer method (about 5%). Since mesh grids were not exposed to a metal etchant solution during our transfer process, the mesh grid material for graphene transfer was not limited. Our graphene transfer method produces high quality and single layer graphene for a graphene TEM grid. Consequently, the formvar assisted graphene transfer method solves disadvantages of polymer support method and direct transfer.