Live-Cell Imaging of MicroRNA Expression via Photoinduced Electron Transfer Controlled by Catalytic Hairpin Assembly.

MicroRNAs (miRNAs) serve as emerging biomarkers for a range of diseases, and their quantitative analysis draws increasing attention. Yet, current invasive methods limit continuous tracking within living cells. To overcome this, a nonenzymatic DNA-based nanoprobe is developed for dynamic, noninvasive miRNA tracking via live-cell imaging. This probe features a unique hairpin DNA structure with five guanines that act as internal quenchers, suppressing fluorescence from an attached fluorophore via photoinduced electron transfer. Target miRNA initiates toehold-mediated strand displacement, restoring, and amplifying the fluorescence signal. Additionally, by introducing a single mismatch to the hairpin DNA, the nanoprobe's sensitivity is significantly enhanced, lowering the detection limit to about 60 pM without compromising specificity. To optimize intracellular delivery for prolonged monitoring, the nanoprobe is encapsulated within multilamellar lipid nanovesicles, fluorescently labeled for dual-wavelength ratiometric analysis. The proposed nanoprobe demonstrates a significant advance in live-cell miRNA detection, promising enhanced in situ analysis for a better understanding of miRNAs' pathophysiological function.

Intranasal delivery of the cytoplasmic domain of CTLA-4 using a novel protein transduction domain prevents allergic inflammation.

CTLA-4 is a negative regulator of T-cell activation, and its inhibitory effects can be accomplished either by competition with CD28 or by transmitting negative signals through its intracellular domain. To utilize the cytoplasmic domain of CTLA-4 to suppress allergic inflammation, we fused it to a novel protein-transduction domain in the human transcriptional factor Hph-1. Transduction efficiency was verified in vitro and in vivo after ocular, intranasal and intradermal administration. After transduction into T cells, the Hph-1-ctCTLA-4 fusion protein inhibited the production of interleukin (IL)-2, and downregulated CD69 and CD25. Intranasal administration of Hph-1-ctCTLA-4 resulted in markedly reduced infiltration of inflammatory cells, secretion of T helper type 2 (T(H)2) cytokines, serum IgE levels and airway hyper-responsiveness in a mouse model of allergic airway inflammation. These results indicated that Hph-1-ctCTLA-4 constitutes an effective immunosuppressive protein drug for potential use in the treatment of allergic asthma, via nasal administration.

Target-Catalyzed Self-Assembly of DNA-Streptavidin Nanogel for Enzyme-Free miRNA Assay.

Rapid, sensitive, specific, and user-friendly microRNA (miRNA) assays are in high demand for point-of-care diagnosis. Target-catalyzed toehold-mediated strand displacement (TMSD) has received increasing attention as an enzyme-free molecular tool for DNA detection. However, the application of TMSD to miRNA targets is challenging because relatively weak DNA/RNA hybridization leads to failure in the subtle kinetic control of multiple hybridization steps. Here, a simple method is presented for miRNA assay based on the one-pot self-assembly of Y-shaped DNAs with streptavidin via an miRNA-catalyzed TMSD cascade reaction. A single miRNA catalyzes the opening cycle of DNA hairpin loops to generate multiple Y-shaped DNAs carrying biotin and a quencher at the end of their arms. Introducing a single base-pair mismatch near the toehold facilitates RNA-triggered strand displacement while barely disturbing nonspecific reactions. The Y-shaped DNAs are self-assembled with fluorescently labeled streptavidin (sAv), which produces nanoscale DNA-sAv nanogel particles mediating efficient Förster resonance energy transfer in their 3D network. The enhancing effect dramatically reduces the detection limit from the nanomolar level to the picomolar level. This work proves that TMSD-based DNA nanogel with a base-pair mismatch incorporated to a hairpin structure is a promising approach towards sensitive and accurate miRNA assay.

Development and validation of the Self-Harm Screening Inventory (SHSI) for adolescents.

Despite the rapidly increasing rate of non-suicidal self-injury (NSSI) among adolescents, there is a dearth of culturally appropriate psychological measures screening for NSSI among the adolescents in the Asian countries. This study aimed to develop and validate the Self-Harm Screening Inventory (SHSI), a culturally sensitive and suitable scale for screening adolescents for NSSI. In total, 514 Korean adolescents (aged 12-16 years) were recruited nationwide. All participants gave informed consent and completed the online self-report measures on NSSI, depression, anxiety, and self-esteem. Thereafter, preliminary items were developed through a series of steps: literature review, ratings of experts on self-harm and suicide, and statistical analyses. Ten of the 20 preliminary items were eliminated after exploratory factor analysis due to low endorsement and factor loading (less than .70). The final version of the SHSI comprised 10 binary items relating to self-harm behaviors within the past year (e.g., cut my body with sharp objects, hit my body). A confirmatory factor analysis supported a one-factor structure, as hypothesized. The one-factor model had a good model fit (x2(35) = 84.958, p < .001, RMSEA = .053, CFI = .981, TLI = .975, SRMR = .124). The SHSI also had good internal consistency (Cronbach's alpha = .795) and 4-week test-retest reliability (r = .786, p < .01). The SHSI had high correlations with another self-harm related scale, the Self-Harm Inventory (r = .773, p < .01), and moderate correlations with the Child Depression Inventory (r = .484, p < .01) and Revised Children's Manifest Anxiety Scale (r = .433, p < .01). Additionally, the SHSI was negatively correlated with the Rosenberg Self-Esteem Scale (r = -.399, p < .01). The findings indicate that the SHSI is a reliable and valid measure for the screening of self-harm behaviors among adolescents.

Clinical and Genetic Characteristics of Korean Patients Diagnosed with Chronic Enteropathy Associated with SLCO2A1 Gene: A KASID Multicenter Study.

Background/Aims: Chronic enteropathy associated with SLCO2A1 gene (CEAS), an inherited disease characterized by nonspecific intestinal ulcers, has emerged in the Japanese population via loss-of-function mutations in the SLCO2A1 gene. We aimed to investigate the clinical and genetic characteristics of Korean patients diagnosed with CEAS. Methods: From July 2018 to July 2021, we performed Sanger sequencing of the SLCO2A1 gene in 46 patients with chronic intestinal ulcers. CEAS was confirmed based on known SLCO2A1 mutations. We summarized the clinical characteristics of patients with confirmed CEAS. Results: Fourteen out of 46 patients (30.4%) had genetically confirmed CEAS, and two SLCO2A1 variants were detected (splicing site variant c.940+1G>A and nonsense mutation [p.R603X] in SLCO2A1). Twelve patients (85.7%) were females and the median age at diagnosis of CEAS was 44.5 years. All patients presented with abdominal pain, and 13 patients (92.9%) presented with anemia (median hemoglobin, 9.6 g/dL). Ten patients (71.4%) had hypoalbuminemia (median, 2.7 g/dL). The most commonly involved site was the ileum (13/14, 92.9%). Manifestations of primary hypertrophic osteoarthropathy (PHO), such as digital clubbing, pachydermia, and periostosis were observed in five patients (28.6%) and two male patients and one female patient satisfied all major PHO diagnostic criteria. Conclusions: The clinical and genetic characteristics of Korean patients with confirmed CEAS were similar to those reported in the literature. CEAS should be considered in the differential diagnosis for patients with unexplained chronic nonspecific ulcers of the small intestine.

Short DNA-catalyzed formation of quantum dot-DNA hydrogel for enzyme-free femtomolar specific DNA assay.

Fast, sensitive, specific, and user-friendly DNA assay is a key technique for the next generation point-of-care molecular diagnosis. However, high-cost, time-consuming, and complicated enzyme-based DNA amplification step is essential to achieve high sensitivity. Herein, a short target DNA-catalyzed formation of quantum dot (QD)-DNA hydrogel is proposed as a new DNA assay platform satisfying the above requirements. A single-stranded target DNA catalyzes the opening cycle of DNA hairpin loops, which are quickly self-assembled with DNA-functionalized QDs to generate QD-DNA hydrogel. The three-dimensional hydrogel network allows efficient resonance energy transfer, dramatically lowering the limit of detection down to ~6 fM without enzymatic DNA amplification. The QD-DNA hydrogel also enables a rapid detection (1 h) with high specificity even for a single-base mismatch. The clinical applicability of the QD-DNA hydrogel is demonstrated for the Klebsiella pneumoniae carbapenemase gene, one of the key targets of drug-resistant pathogenic bacteria.

The outcome of ultrasound-guided needle decompression and steroid injection in calcific tendinitis.

HYPOTHESIS: Needle lavage is frequently performed before consideration of surgical removal in shoulders with calcific tendinitis because this may avoid surgery. However, its role in nonoperative treatment has not been fully investigated in terms of clinical and radiographic response. We hypothesized that needle decompression and subacromial steroid injection would show good clinical results in chronic calcific tendinitis patients. MATERIALS AND METHODS: Thirty-five shoulders in 30 consecutive patients with painful calcific tendinitis were treated by ultrasound-guided needle decompression and subacromial corticosteroid injection. Patients were prospectively evaluated using American Shoulder and Elbow Surgeons (ASES) and Constant scores at 1, 3, and 6 months after the intervention. Size and morphology of the calcific deposits were compared with those in baseline radiographs at each visit. RESULTS: At 6 months after the index procedure, 25 shoulders (71.4%) showed ASES and Constant score improvements from 48.0 and 53.7 to 84.6 and 87.9, respectively (P < .01). Ten shoulders (28.6%) showed no symptom relief at the last follow-up. In shoulders with pain improvement, the mean size of calcific deposits reduced from 13.6 to 5.6 mm (P < .01), and in shoulders with no pain improvement or that underwent operation, mean size was 13.1 mm at initial visits and 12.7 mm at final visits (P = .75). DISCUSSION: Shoulders showing little evidence of deposit size reduction at 6 months after needle decompression are less likely to achieve symptomatic improvement and may be considered as candidates for surgical removal. CONCLUSION: Needle decompression with subacromial steroid injection is effective in 71.4% of calcific tendinitis within 6 months. The size of calcific deposits in patients that achieved symptom relief was reduced.

The condylar cutoff sign: quantifying lateral femoral condylar hypoplasia in a complete discoid meniscus.

UNLABELLED: We describe the condylar cutoff sign, a radiographic sign in knees with a discoid lateral meniscus, for diagnosis of complete discoid meniscus. We retrospectively reviewed tunnel-view radiographs of 100 patients (100) knees including 50 with complete discoid lateral meniscus and 50 with normal menisci. All patients were 18 years of age or older. All of the knees were arthroscopically examined. We developed a method to measure the prominence of the femoral condyle adjacent to the intercondylar notch on a tunnel view of the knee. The prominence ratio, the ratio of the medial and lateral condylar prominence, was compared and analyzed. The measurements were performed by three observers on two separate occasions to determine reliability. The intraobserver and interobserver variability study revealed high reliability with correlation coefficients ranging from 0.86 to 0.99. Using a cutoff ratio of 0.8, we observed a major difference between the two groups and found the ratio had 76% sensitivity, 96% specificity, 95% positive predictive value, and 80% negative predictive value in every set of measurements. The sign has high specificity and reliability but might not apply to younger patients. LEVEL OF EVIDENCE: Level II, diagnostic study. See the Guidelines for Authors for a complete description of levels of evidence.

Treatment of periprosthetic hip infection caused by resistant microorganisms using 2-stage reimplantation protocol.

To determine the effectiveness of staged reimplantation for the treatment of periprosthetic hip infection caused by resistant microorganisms, we performed a retrospective case-control study on 37 consecutive patients who had a culture-proven periprosthetic hip infection treated using a 2-stage reimplantation protocol. Twenty-four patients infected by resistant microorganisms were compared with 13 patients infected by nonresistant microorganisms. The second-stage reimplantation procedure was possible in 34 (92%) of the 37 patients, but the remaining 3 required permanent resection because of persistent infection. At a mean follow-up of 4.4 years, there were 4 recurrent infections and 1 aseptic cup loosening. Overall treatment failure rate was 22%. All failures occurred only in the resistant microorganism group and none in the nonresistant microorganism group (33% vs 0%; P = .032). Current 2-stage reimplantation protocol showed a high rate of treatment failure in our patients with a periprosthetic hip infection caused by resistant microorganisms.

Subnanomolar FRET-Based DNA Assay Using Thermally Stable Phosphorothioated DNA-Functionalized Quantum Dots.

Quantum dots (QDs) can serve as an attractive Förster resonance energy transfer (FRET) donor for DNA assay due to their excellent optical properties. However, the specificity and sensitivity of QD-based FRET analysis are prominently reduced by nonspecific DNA adsorption and poor colloidal stability during DNA hybridization, which hinders the practical applications of QDs as a biosensing platform. Here, we report subnanomolar FRET assay of DNA through the stabilization of DNA/QD interface using DNA-functionalized QDs with phosphorothioated single-stranded DNA (pt-ssDNA) as a multivalent ligand in an aqueous solution. In situ DNA functionalization was achieved during the aqueous synthesis of CdTe/CdS QDs, resulting in the maximum photoluminescence quantum yields of 76.9% at an emission wavelength of 732 nm. Conventional monothiolated ssDNA-capped QDs exhibited particle aggregation and photoluminescence (PL) quenching during DNA hybridization at 70 °C due to the dissociation of surface ligands. Such colloidal instability induced the nonspecific adsorption of DNA, resulting in false-positive signal and decreased sensitivity with the limit of detection (LOD) of 16.1 nM. In contrast, the pt-ssDNA-functionalized QDs maintained their colloidal stability and PL properties at elevated temperatures. The LOD of the pt-ssDNA-functionalized QDs was >30 times lower (0.47 nM) while maintaining the high specificity to a target sequence because the strong multivalent binding of pt-ssDNA to the surface of QDs prevents the detachment of pt-ssDNA and nonspecific adsorption of DNA. The study suggests that the ligand design to stabilize the surface of QDs in an aqueous milieu is critically important for the high performance of QDs for specific DNA assay.

Paclitaxel-induced formation of 3D nanocrystal superlattices within injectable protein-based hybrid nanoparticles.

Self-assembly of monodisperse superparamagnetic iron oxide nanocrystals into a close-packed, three-dimensional (3D) superlattice is designed within cross-linked protein-based nanoparticles composed of human serum albumin and polyethylene glycol. The prepared nanoparticles are very stable in serum and exhibit a high T2 relaxivity as well as anti-cancer activity, indicating the practical benefits of ordering nanocrystals.

Protein-quantum dot nanohybrids for bioanalytical applications.

Quantum dots (QDs) coupled with biomolecules play an important role as optically and chemically stable bioimaging agents for various applications. These inorganic-biological hybrid conjugates have been demonstrated as powerful fluorescence tools for sensing, diagnostics, and labeling. This review focuses on protein-QD nanohybrids for different types of bioanalytical applications. There are various strategies to modify the surface properties of QDs to produce protein-QD nanohybrids that are stable in biological fluids. We expect that multifunctional protein-QD nanohybrids can be used as a powerful optical probe for various biological applications. WIREs Nanomed Nanobiotechnol 2016, 8:178-190. doi: 10.1002/wnan.1345 For further resources related to this article, please visit the WIREs website.

Controlling surface defects of non-stoichiometric copper-indium-sulfide quantum dots.

Quantum dots (QDs) can be used for a wide range of practical applications including solar energy conversion, light-emitting display, bio-imaging, and sensing. However, toxic heavy metal elements of Pb- and Cd-based QDs cause potential environmental problems and limit their wide applicability. To overcome this limitation, CuInS2 (CIS) QDs, which have a bulk bandgap energy of 1.5eV and relatively high absorptivity, can be a good alternative. However the photoluminescence quantum yield (PLQY) of CIS QDs is too low for practical applications. Here we investigate the effects of experimental factors in the solution synthesis of CIS/ZnS QDs on intrinsic defects and surface defects from photoluminescence (PL) analysis. A heating-up method is used with dodecanethiol as a sulfur source, a ligand, and a medium. The Cu-to-In feeding ratio is changed to control the PL spectrum in the range of visible to near infrared (NIR) frequencies. The PLQY is increased above 40% in all of the ranges through ZnS shell passivation and additional process optimization (e.g., controlled cooling rate and additional feeding of In(3+) ion precursor). This work demonstrates the role of intrinsic defects in PL and the importance of suppressing the formation of the surface defects to increase the PLQY.

Small-dose-sensitive X-ray image pixel with HgI2 photoconductor and amorphous oxide thin-film transistor.

A new X-ray image sensor is demonstrated with an oxide thin-film transistor backplane and HgI2 sensing material. It displays outstanding image quality under a low X-ray exposure and a low electric field. It is promising as a state-of-the-art device to realize highly resolved images at a low X-ray dose for a variety of medical X-ray imaging applications.

Painful jerk test: a predictor of success in nonoperative treatment of posteroinferior instability of the shoulder.

BACKGROUND: The jerk test has been used as a diagnostic test of the posteroinferior instability of the shoulder. Pain may or may not be associated with posterior clunking during the jerk test. PURPOSE: To evaluate the presence or absence of pain with the jerk test as a predictor of the success of nonoperative treatment for posteroinferior instability of the shoulder and to identify the pathologic lesion responsible for the pain in the jerk test. STUDY DESIGN: Retrospective review of prospectively collected data. METHODS: Eighty-one patients (89 shoulders) who had posteroinferior instability with a positive posterior clunk in their shoulders during the jerk test were nonoperatively treated. The patients were divided into 2 groups with respect to the presence of pain in the jerk test: the painless jerk group (54 shoulders) and the painful jerk group (35 shoulders). Response to the nonoperative treatment was evaluated after at least a 6-month rehabilitation program. Patients who did not respond to the rehabilitation underwent arthroscopic examination to identify any pathologic lesions. RESULTS: The painful jerk group had a higher failure rate with nonoperative treatment (P < .001). In the painless jerk group, 50 shoulders (93%) responded to the rehabilitation program after a mean of 4 months. Four shoulders (7%) were unresponsive to the rehabilitation. In the painful jerk group, 5 shoulders (16%) were successful with the rehabilitation, whereas the other 30 shoulders (84%) failed. All 34 shoulders that were unresponsive to the rehabilitation had a variable degree of posteroinferior labral lesions. CONCLUSIONS: The jerk test is a hallmark for predicting the prognosis of nonoperative treatment for posteroinferior instability. Shoulders with symptomatic posteroinferior instability and a painful jerk test have posteroinferior labral lesions.

Self-aligned top-gate amorphous indium zinc oxide thin-film transistors exceeding low-temperature poly-Si transistor performance.

Thin-film transistor (TFT) is a key component of active-matrix flat-panel displays (AMFPDs). These days, the low-temperature poly silicon (LTPS) TFTs are to match with advanced AMFPDs such as the active matrix organic light-emitting diode (AMOLED) display, because of their high mobility for fast pixel switching. However, the manufacturing process of LTPS TFT is quite complicated, costly, and scale-limited. Amorphous oxide semiconductor (AOS) TFT technology is another candidate, which is as simple as that of conventioanl amorphous (a)-Si TFTs in fabrication but provides much superior device performances to those of a-Si TFTs. Hence, various AOSs have been compared with LTPS for active channel layer of the advanced TFTs, but have always been found to be relatively inferior to LTPS. In the present work, we clear the persistent inferiority, innovating the device performaces of a-IZO TFT by adopting a self-aligned coplanar top-gate structure and modifying the surface of a-IZO material. Herein, we demonstrate a high-performance simple-processed a-IZO TFT with mobility of ∼157 cm(2) V(-1) s(-1), SS of ∼190 mV dec(-1), and good bias/photostabilities, which overall surpass the performances of high-cost LTPS TFTs.

High-performance low-cost back-channel-etch amorphous gallium-indium-zinc oxide thin-film transistors by curing and passivation of the damaged back channel.

High-performance, low-cost amorphous gallium-indium-zinc oxide (a-GIZO) thin-film-transistor (TFT) technology is required for the next generation of active-matrix organic light-emitting diodes. A back-channel-etch structure is the most appropriate device structure for high-performance, low-cost a-GIZO TFT technology. However, channel damage due to source/drain etching and passivation-layer deposition has been a critical issue. To solve this problem, the present work focuses on overall back-channel processes, such as back-channel N2O plasma treatment, SiOx passivation deposition, and final thermal annealing. This work has revealed the dependence of a-GIZO TFT characteristics on the N2O plasma radio-frequency (RF) power and frequency, the SiH4 flow rate in the SiOx deposition process, and the final annealing temperature. On the basis of these results, a high-performance a-GIZO TFT with a field-effect mobility of 35.7 cm(2) V(-1) s(-1), a subthreshold swing of 185 mV dec(-1), a switching ratio exceeding 10(7), and a satisfactory reliability was successfully fabricated. The technology developed in this work can be realized using the existing facilities of active-matrix liquid-crystal display industries.

A Broken Drill-bit Fragment Causing Severe Radiating Pain after Cervical Total Disc Replacement: A Case Report.

This is a case report of a 38-year-old man with severe radiating pain on upper extremity after cervical total disc replacement (TDR). We faced an unusual complication that has not been reported yet. He underwent cervical TDR for left central disc protrusion on C5-6. After the surgery, preoperative symptom disappeared. However, at postoperative 1 year, he complained severe right-sided radiating pain that had a sudden onset. On postoperative X-ray, a metal fragment which seemed like a broken drill bit was shown within the spinal canal. To remove that, right-sided anterior microforaminotomy on C5-6 was performed and the metal fragment was removed successfully. After that, anterior fusion was done because the motion of the artificial disc was minimal and the removed structure seemed to attenuate stability during cervical motion. The operation resulted in prompt symptomatic relief. During cervical TDR, particular attention should be paid to the procedures that require using drill-bits.