Impaired binding affinity of YTHDC1 with METTL3/METTL14 results in R-loop accumulation in myelodysplastic neoplasms with DDX41 mutation.

DEAD box helicase 41 (DDX41) mutations are the most prevalent predisposition to familial myelodysplastic syndrome (MDS). However, the precise roles of these variants in the pathogenesis of MDS have yet to be elucidated. Here, we discovered a novel mechanism by which DDX41 contributes to R-loop-induced DNA damage responses (DDR) in cooperation with the m6A-METTL complex (MAC) and YTHDC1 using DDX41 knockout (KO) and DDX41 knock-in (KI, R525H, Y259C) cell lines as well as primary samples from MDS patients. Compared to wild type (WT), DDX41 KO and KI led to increased levels of m6A RNA methylated R-loop. Interestingly, we found that DDX41 regulates m6A/R-loop levels by interacting with MAC components. Further, DDX41 promoted the recruitment of YTHDC1 to R-loops by promoting the binding between METTL3 and YTHDC1, which was dysregulated in DDX41-deficient cells, contributing to genomic instability. Collectively, we demonstrated that DDX41 plays a key role in the physiological control of R-loops in cooperation with MAC and YTHDC1. These findings provide novel insights into how defects in DDX41 influence MDS pathogenesis and suggest potential therapeutic targets for the treatment of MDS.

3D-cultured blastoids model human embryogenesis from pre-implantation to early gastrulation stages.

Naive human pluripotent stem cells have the remarkable ability to self-organize into blastocyst-like structures ("blastoids") that model lineage segregation in the pre-implantation embryo. However, the extent to which blastoids can recapitulate the defining features of human post-implantation development remains unexplored. Here, we report that blastoids cultured on thick three-dimensional (3D) extracellular matrices capture hallmarks of early post-implantation development, including epiblast lumenogenesis, rapid expansion and diversification of trophoblast lineages, and robust invasion of extravillous trophoblast cells by day 14. Extended blastoid culture results in the localized activation of primitive streak marker TBXT and the emergence of embryonic germ layers by day 21. We also show that the modulation of WNT signaling alters the balance between epiblast and trophoblast fates in post-implantation blastoids. This work demonstrates that 3D-cultured blastoids offer a continuous and integrated in vitro model system of human embryonic and extraembryonic development from pre-implantation to early gastrulation stages.

Dual-modality imaging system for monitoring human heart organoids beating in vitro.

To reveal the three-dimensional microstructure and calcium dynamics of human heart organoids (hHOs), we developed a dual-modality imaging system combining the advantages of optical coherence tomography (OCT) and fluorescence microscopy. OCT provides high-resolution volumetric structural information, while fluorescence imaging indicates the electrophysiology of the hHOs' beating behavior. We verified that concurrent OCT motion mode (M-mode) and calcium imaging retrieved the same beating pattern from the heart organoids. We further applied dynamic contrast OCT (DyC-OCT) analysis to strengthen the verification and localize the beating clusters inside the hHOs. This imaging platform provides a powerful tool for studying and assessing hHOs in vitro, with potential applications in disease modeling and drug screening.

Alteration of replication protein A binding mode on single-stranded DNA by NSMF potentiates RPA phosphorylation by ATR kinase.

Replication protein A (RPA), a eukaryotic single-stranded DNA (ssDNA) binding protein, dynamically interacts with ssDNA in different binding modes and plays essential roles in DNA metabolism such as replication, repair, and recombination. RPA accumulation on ssDNA due to replication stress triggers the DNA damage response (DDR) by activating the ataxia telangiectasia and RAD3-related (ATR) kinase, which phosphorylates itself and downstream DDR factors, including RPA. We recently reported that the N-methyl-D-aspartate receptor synaptonuclear signaling and neuronal migration factor (NSMF), a neuronal protein associated with Kallmann syndrome, promotes RPA32 phosphorylation via ATR upon replication stress. However, how NSMF enhances ATR-mediated RPA32 phosphorylation remains elusive. Here, we demonstrate that NSMF colocalizes and physically interacts with RPA at DNA damage sites in vivo and in vitro. Using purified RPA and NSMF in biochemical and single-molecule assays, we find that NSMF selectively displaces RPA in the more weakly bound 8- and 20-nucleotide binding modes from ssDNA, allowing the retention of more stable RPA molecules in the 30-nt binding mode. The 30-nt binding mode of RPA enhances RPA32 phosphorylation by ATR, and phosphorylated RPA becomes stabilized on ssDNA. Our findings provide new mechanistic insight into how NSMF facilitates the role of RPA in the ATR pathway.

ZNF212 promotes genomic integrity through direct interaction with TRAIP.

TRAIP is a key factor involved in the DNA damage response (DDR), homologous recombination (HR) and DNA interstrand crosslink (ICL) repair. However, the exact functions of TRAIP in these processes in mammalian cells are not fully understood. Here we identify the zinc finger protein 212, ZNF212, as a novel binding partner for TRAIP and find that ZNF212 colocalizes with sites of DNA damage. The recruitment of TRAIP or ZNF212 to sites of DNA damage is mutually interdependent. We show that depletion of ZNF212 causes defects in the DDR and HR-mediated repair in a manner epistatic to TRAIP. In addition, an epistatic analysis of Zfp212, the mouse homolog of human ZNF212, in mouse embryonic stem cells (mESCs), shows that it appears to act upstream of both the Neil3 and Fanconi anemia (FA) pathways of ICLs repair. We find that human ZNF212 interacted directly with NEIL3 and promotes its recruitment to ICL lesions. Collectively, our findings identify ZNF212 as a new factor involved in the DDR, HR-mediated repair and ICL repair though direct interaction with TRAIP.

First report of alstroemeria mosaic virus infecting Alstroemeria in Korea.

Alstroemeria, a member of the Alstroemriaceae family, is a popular cut flower plant with a long-base life and a wide variety of flower colors. It is widely cultivated in many countries, especially in Central and South America. However, numerous viruses such as alstroemeria carlavirus (AlCV), alstroemeria mosaic virus (AlMV), cucumber mosaic virus (CMV), tomato spotted wilt virus (TSWV), alstroemeria streak virus (AlSV), and impatiens necrotic virus (INSV) can infect Alstroemeria and significantly decrease its yield (Kim, 2020). Among these viruses, AlMV is well known to cause an endemic viral disease in the Netherlands (Corine M. et al. 1992). AlMV is a member of the genus potyvirus in the family Potyviridae, one of the most widely distributed families of plant viruses. In 2021, symptomatic alstroemeria plants showing interveinal leaf streaking with elongated light green and chlorosis of leaves were identified from farms in a greenhouse in Gwangju, South Korea. Potyvirus-like particles (approximately 750-800 nm in length) were observed from sap of the symptomatic plants by electron microscope (Supplementary Fig. 1). To confirm virus infection, total RNA was extracted from an alstroemeria leaf using a Beniprep® Super Plant RNA extraction kit (IVT7005, Invirustech Co., Korea). A cDNA library was synthesized and analyzed by high throughput sequencing (HTS) using an Illumina NovaSeq6000 S4 sequencer. A total of 48,072,240 raw reads were obtained after quality filtering with FastQC. Remaining sequences were de novo assembled into contigs with a Trinity assembler. Nucleotide blast analysis of contigs against NCBI viral reference database revealed that 24 assembled contigs (> 1,000 bp) were sequences of AlMV. To confirm AlMV detection, raw reads were mapped to known AlMV complete genome (9,774 bp) using Bowtie2 program. Results showed that a total of 4,698,112 reads were mapped. A consensus sequence (9,778 bp, accession no. LC709275) was then obtained. To verify the presence of AlMV, RT-PCR assay was conducted with AlMV's CP gene-specific primers: AlMV-F (5'-CACGAGGCTGTGAAACAAGC -3') and AlMV-R (5'- CCAGGCGACACGGCTAAATA-3'). PCR products of the expected size (538 bp) were cloned, sequenced, and subjected to GenBank BLASTn search. A 538 bp partial CP sequence was used for BLAST analysis which revealed that it shared 100% identities with the consensus sequence (LC709275) and 96.99~98.76% nucleotide identities with four AlMV isolates (MK440140, NC043135, MT892648, DQ295032). Phylogenetic analysis based on partial CP sequences of representative members of potyviruses (family Potyviridae) using 1,000 bootstrap replicates based on either neighbor-joining or Kimura 2 parameter methods in MEGA-X revealed that AlMV isolate JNU-2 was grouped together with the four known AlMV isolates (Supplementary Fig. 2). To determine the incidence of AlMV in a greenhouse, 30 alstroemeria samples were collected and tested by RT-PCR. Results showed that 23 samples were positive for AlMV by PCR-gel electrophoresis and Sanger sequencing, suggesting a high incidence of AlMV infection. To the best of our knowledge, this is the first report of natural infection with AlMV in Alstroemeria in Korea. Further surveys of AlMV infection in greenhouses will help us prevent the spread of this viral disease in Alstroemeria.

Spinal orexin A attenuates opioid-induced mechanical hypersensitivity in the rat.

Background: Repeated administration of opioid analgesics for pain treatment can produce paradoxical hyperalgesia via peripheral and/or central mechanisms. Thus, this study investigated whether spinally (centrally) administered orexin A attenuates opioid-induced hyperalgesia (OIH). Methods: [D-Ala2, N-Me-Phe4, Gly5-ol]-enkephalin (DAMGO), a selective µ-opioid receptor agonist, was used to induce mechanical hypersensitivity and was administered intradermally (4 times, 1-hour intervals) on the rat hind paw dorsum. To determine whether post- or pretreatments with spinal orexin A, dynorphin A, and anti-dynorphin A were effective in OIH, the drugs were injected through an intrathecal catheter whose tip was positioned dorsally at the L3 segment of the spinal cord (5 µg for all). Mechanical hypersensitivity was assessed using von Frey monofilaments. Results: Repeated intradermal injections of DAMGO resulted in mechanical hypersensitivity in rats, lasting more than 8 days. Although the first intrathecal treatment of orexin A on the 6th day after DAMGO exposure did not show any significant effect on the mechanical threshold, the second (on the 8th day) significantly attenuated the DAMGO-induced mechanical hypersensitivity, which disappeared when the type 1 orexin receptor (OX1R) was blocked. However, intrathecal administration of dynorphin or an anti-dynorphin antibody (dynorphin antagonists) had no effect on DAMGO-induced hypersensitivity. Lastly, pretreatment with orexin A, dynorphin, or anti-dynorphin did not prevent DAMGO-induced mechanical hypersensitivity. Conclusions: Spinal orexin A attenuates mechanical hyperalgesia induced by repetitive intradermal injections of DAMGO through OX1R. These data suggest that OIH can be potentially treated by activating the orexin A-OX1R pathway in the spinal dorsal horn.

Reciprocal interactions among Cobll1, PACSIN2, and SH3BP1 regulate drug resistance in chronic myeloid leukemia.

Cobll1 affects blast crisis (BC) progression and tyrosine kinase inhibitor (TKI) resistance in chronic myeloid leukemia (CML). PACSIN2, a novel Cobll1 binding protein, activates TKI-induced apoptosis in K562 cells, and this activation is suppressed by Cobll1 through the interaction between PACSIN2 and Cobll1. PACSIN2 also binds and inhibits SH3BP1 which activates the downstream Rac1 pathway and induces TKI resistance. PACSIN2 competitively interacts with Cobll1 or SH3BP1 with a higher affinity for Cobll1. Cobll1 preferentially binds to PACSIN2, releasing SH3BP1 to promote the SH3BP1/Rac1 pathway and suppress TKI-mediated apoptosis and eventually leading to TKI resistance. Similar interactions among Cobll1, PACSIN2, and SH3BP1 control hematopoiesis during vertebrate embryogenesis. Clinical analysis showed that most patients with CML have Cobll1 and SH3BP1 expression at the BC phase and BC patients with Cobll1 and SH3BP1 expression showed severe progression with a higher blast percentage than those without any Cobll1, PACSIN2, or SH3BP1 expression. Our study details the molecular mechanism of the Cobll1/PACSIN2/SH3BP1 pathway in regulating drug resistance and BC progression in CML.

Effect of Iron Content on Corrosion Properties of Pure Titanium as Grain Refiner.

Microstructures and corrosion properties of pure titanium were characterized when iron was used as a grain refiner. The added Fe element acted as a strong grain refiner for pure titanium by forming ß Ti phase at grain boundaries, and 0.15 wt% Fe was revealed to be a sufficient amount to make the grain size of pure titanium below 20 µm, which was the requirement for the desired titanium cathode. However, corrosion resistance was decreased with the Fe amount added. From the open circuit potential (OCP) results, it was obvious that the TiO2 stability against the reducing acid environment was deteriorated with the Fe amount, which seemed to be the main reason for the decreased corrosion resistance. Electrochemical impedance spectroscopy (EIS) results showed that both the decrease in the compact oxide film's resistance (Rb) and the appearance of the outer porous film occurred as a result of the dissolution of the TiO2 layer, whose phenomena became more apparent as more Fe was added.

RNF126 is a positive regulator of TRAF3 ubiquitination.

Ubiquitination and deubiquitination of signaling molecules are critical regulatory mechanisms in various biological contexts such as inflammatory signaling and the DNA damage response. Thus, finely tuned regulation of protein ubiquitination is essential for maintaining cellular homeostasis. Here, we showed that the RING finger protein RNF126 interacts with TRAF3 and promotes its K63-linked polyubiquitination, which is a crucial step in the TRAF3-dependent antiviral response. We found that RNF126 also interacts with OTUB1, a deubiquitinating enzyme that negatively regulates K63-linked ubiquitination of TRAF3. RNF126 promotes ubiquitination of OTUB1, leading to reduced deubiquitinating activity toward TRAF3. Moreover, RNF126 promotes ubiquitination of OTUB1 on cysteine 91, which is reportedly required for its catalytic activity. Taken together, our results suggest that RNF126 positively regulates the antiviral response by directly promoting K63-linked polyubiquitination of TRAF3 and by reducing OTUB1 activity.

Voltage-dependent calcium channel ß subunit-derived peptides reduce excitatory neurotransmission and arterial blood pressure.

AIMS: Voltage-dependent calcium channels (VDCCs) play an important role in various physiological functions in the nervous system and the cardiovascular system. In L-, N-, P/Q-, and R-type VDCCs, ß subunit assists the channels for membrane targeting and modulates channel properties. In this study, we investigated whether an inhibition of the ß subunit binding to α subunit, the pore-forming main subunit of VDCCs, have any effect on channel activation and physiological functions. MAIN METHODS: Peptides derived from the specific regions of ß subunit that bind to the α-interaction domain in I-II linker of α subunit were manufactured, presuming that the peptides interrupt α-ß subunit interaction in the channel complex. Then, they were tested on voltage-activated Ca2+ currents recorded in acutely isolated trigeminal ganglion (TG) neurons, excitatory postsynaptic currents (EPSCs) in the spinal dorsal horn neurons, and arterial blood pressure (BP) recorded from the rat femoral artery. KEY FINDINGS: When applied internally through patch pipettes, the peptides decreased the peak amplitudes of the voltage-activated Ca2+ currents. After fusing with HIV transactivator of transcription (TAT) sequence to penetrate cell membrane, the peptides significantly decreased the peak amplitudes of Ca2+ currents and the peak amplitudes of EPSCs upon the external application through bath solution. Furthermore, the TAT-fused peptides dose dependently reduced the rat BP when administered intravenously. SIGNIFICANCE: These data suggest that an interruption of α-ß subunit association in VDCC complex inhibits channel activation, thereby reducing VDCC-mediated physiological functions such as excitatory neurotransmission and arterial BP.

Serial optical coherence microscopy for label-free volumetric histopathology.

The observation of histopathology using optical microscope is an essential procedure for examination of tissue biopsies or surgically excised specimens in biological and clinical laboratories. However, slide-based microscopic pathology is not suitable for visualizing the large-scale tissue and native 3D organ structure due to its sampling limitation and shallow imaging depth. Here, we demonstrate serial optical coherence microscopy (SOCM) technique that offers label-free, high-throughput, and large-volume imaging of ex vivo mouse organs. A 3D histopathology of whole mouse brain and kidney including blood vessel structure is reconstructed by deep tissue optical imaging in serial sectioning techniques. Our results demonstrate that SOCM has unique advantages as it can visualize both native 3D structures and quantitative regional volume without introduction of any contrast agents.

Quantitative Screening of Cervical Cancers for Low-Resource Settings: Pilot Study of Smartphone-Based Endoscopic Visual Inspection After Acetic Acid Using Machine Learning Techniques.

BACKGROUND: Approximately 90% of global cervical cancer (CC) is mostly found in low- and middle-income countries. In most cases, CC can be detected early through routine screening programs, including a cytology-based test. However, it is logistically difficult to offer this program in low-resource settings due to limited resources and infrastructure, and few trained experts. A visual inspection following the application of acetic acid (VIA) has been widely promoted and is routinely recommended as a viable form of CC screening in resource-constrained countries. Digital images of the cervix have been acquired during VIA procedure with better quality assurance and visualization, leading to higher diagnostic accuracy and reduction of the variability of detection rate. However, a colposcope is bulky, expensive, electricity-dependent, and needs routine maintenance, and to confirm the grade of abnormality through its images, a specialist must be present. Recently, smartphone-based imaging systems have made a significant impact on the practice of medicine by offering a cost-effective, rapid, and noninvasive method of evaluation. Furthermore, computer-aided analyses, including image processing-based methods and machine learning techniques, have also shown great potential for a high impact on medicinal evaluations. OBJECTIVE: In this study, we demonstrate a new quantitative CC screening technique and implement a machine learning algorithm for smartphone-based endoscopic VIA. We also evaluated the diagnostic performance and practicability of the approach based on the results compared to the gold standard and from physicians' interpretation. METHODS: A smartphone-based endoscope system was developed and applied to the VIA screening. A total of 20 patients were recruited for this study to evaluate the system. Overall, five were healthy, and 15 were patients who had shown a low to high grade of cervical intraepithelial neoplasia (CIN) from both colposcopy and cytology tests. Endoscopic VIA images were obtained before a loop electrosurgical excision procedure for patients with abnormal tissues, and their histology tissues were collected. Endoscopic VIA images were assessed by four expert physicians relative to the gold standard of histopathology. Also, VIA features were extracted from multiple steps of image processing techniques to find the differences between abnormal (CIN2+) and normal (≤CIN1). By using the extracted features, the performance of different machine learning classifiers, such as k-nearest neighbors (KNN), support vector machine, and decision tree (DT), were compared to find the best algorithm for VIA. After determining the best performing classifying model, it was used to evaluate the screening performance of VIA. RESULTS: An average accuracy of 78%, with a Cohen kappa of 0.571, was observed for the evaluation of the system by four physicians. Through image processing, 240 sliced images were obtained from the cervicogram at each clock position, and five features of VIA were extracted. Among the three models, KNN showed the best performance for finding VIA within holdout 10-fold cross-validation, with an accuracy of 78.3%, area under the curve of 0.807, a specificity of 80.3%, and a sensitivity of 75.0%, respectively. The trained model performed using an unprovided data set resulted in an accuracy of 80.8%, specificity of 84.1%, and sensitivity of 71.9%. Predictions were visualized with intuitive color labels, indicating the normal/abnormal tissue using a circular clock-type segmentation. Calculating the overlapped abnormal tissues between the gold standard and predicted value, the KNN model overperformed the average assessments of physicians for finding VIA. CONCLUSIONS: We explored the potential of the smartphone-based endoscopic VIA as an evaluation technique and used the cervicogram to evaluate normal/abnormal tissue using machine learning techniques. The results of this study demonstrate its potential as a screening tool in low-resource settings.

Digital subtraction angiography vs. real-time fluoroscopy for detection of intravascular injection during transforaminal epidural block.

Background: Transforaminal epidural block (TFEB) is an effective treatment option for radicular pain. To reduce complications from intravascular injection during TFEB, use of imaging modalities such as real-time fluoroscopy (RTF) or digital subtraction angiography (DSA) has been recommended. In this study, we investigated whether DSA improved the detection of intravascular injection during TFEB at the whole spine level compared to RTF. Methods: We prospectively examined 316 patients who underwent TFEB. After confirmation of final needle position using biplanar fluoroscopy, 2 mL of nonionic contrast medium was injected at a rate of 0.5 mL/s under RTF; 30 s later, 2 mL of nonionic contrast medium was injected at a rate of 0.5 mL/s under DSA. Results: Thirty-six intravascular injections were detected for an overall rate of 11.4% using RTF, with 45 detected for a rate of 14.2% using DSA. The detection rate using DSA was statistically different from that using RTF (p=0.004). DSA detected a significantly higher proportion of intravascular injections at the cervical level than at the thoracic (p=0.009) and lumbar (p=0.011) levels. Conclusion: During TFEB at the whole spine level, DSA was better than RTF for the detection of intravascular injection. Special attention is advised for cervical TFEB, because of a significantly higher intravascular injection rate at this level than at other levels.

Variation of Mechanical Characteristics of Polyurethane Foam: Effect of Test Method.

Polyurethane foam (PUF), a representative insulation material, not only prevents heat conduction but can also support a load. Particular interest in rigid PUF proliferated over the past several years in fields where extreme environments are applied. A closed-cell structure which forms the interior of rigid PUF serves to maximize the utilization of these polymeric foams. Rigid PUF is more sensitive to external conditions such as temperature or restraint than other structural materials such as steel. Depending on the market trends in which utilization of a cryogenic environment is expanding, the tendency of material behavior resulting from the binding effect also needs to be investigated. However, most conventional compression test method standards applicable to rigid PUF do not adequately reflect the restraints. Therefore, this study proposes a method for evaluating the mechanical performance of materials in a more reliable manner than that of conventional tests. Experimental observation and analysis validated this compression evaluation method in which constraints are considered. Consequently, the compressive strength of rigid PUF compared to the results of the conventional test showed a difference of up to 0.47 MPa (approximately 23%) at cryogenic temperatures. This result suggests that there are important factors to consider when assessing performance from a material perspective in an environment where rigid PUF insulation is utilized. It is believed that the test methods newly proposed in this study will provide an experimental framework that can be applied to the evaluation criteria of material properties and reflected in structural design.

GCA links TRAF6-ULK1-dependent autophagy activation in resistant chronic myeloid leukemia.

Imatinib is the first molecularly targeted compound for chronic myeloid leukemia (CML) capable to inhibit BCR-ABL kinase activity. However, recent clinical evidence indicates that a substantial proportion of CML patients exhibit BCR-ABL-dependent or independent resistance to imatinib. Despite the importance of imatinib resistance in CML, the underlying molecular mechanisms of this resistance are largely unknown. Here, we identified GCA (grancalcin) as a critical regulator of imatinib resistance in chronic phase CML via activation of autophagy. Mechanistically, we demonstrated that GCA activates TRAF6 ubiquitin ligase activity to induce Lys63 ubiquitination of ULK1, a crucial regulator of autophagy, resulting in its stabilization and activation. We also highlighted the role of GCA-TRAF6-ULK1 autophagy regulatory axis in imatinib resistance. Our findings represent the basis for novel therapeutic strategies against CML.Abbreviation: ACTB/ß-actin: actin beta; ADM: adrenomedullin; AMBRA1: autophagy and beclin 1 regulator 1; AMPK: AMP-activated protein kinase; ANXA5: annexin A5; CP: cytogenetic response; CML: chronic myeloid leukemia; CUL3: cullin 3; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GCA: grancalcin; Dx: at diagnosis; E-64-d: (2S,3S)-trans-Epoxysuccinyl-L-leucylamido-3-methylbutane ethyl ester; IMres: Imatinib resistance; KLHL20: Kelch-like protein 20; LRMP: lymphoid-restricted membrane protein; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; MMR: major molecular response; NH4Cl: ammonium chloride; PBMCs: peripheral blood mononuclear cells; PTPRC: protein tyrosine phosphatase, receptor type, C; SQSTM1/p62: sequestosome 1; SYK: spleen associated tyrosine kinase; TAP1: transporter 1, ATP binding cassette subfamily B member; TKIs: ABL-specific tyrosine kinase inhibitors; TLR9: toll- like receptor 9; TRAF6: TNF receptor associated factor 6; ULK1: unc-51 like autophagy activating kinase 1.

In Vivo Vibration Measurement of Middle Ear Structure Using Doppler Optical Coherence Tomography: Preliminary Study.

OBJECTIVES: Doppler optical coherence tomography (DOCT) is useful for both, the spatially resolved measurement of the tympanic membrane (TM) oscillation and high-resolution imaging. We demonstrated a new technique capable of providing real-time two-dimensional Doppler OCT image of rapidly oscillatory latex mini-drum and in vivo rat TM and ossicles. METHODS: Using DOCT system, the oscillation of sample was measured at frequency range of 1-4 kHz at an output of 15 W. After the sensitivity of the DOCT system was verified using a latex mini-drum consisting of a 100 µm-thick latex membrane, changes in displacement of the umbo and contacted area between TM and malleus in normal and pathologic conditions. RESULTS: The oscillation cycles of the mini-drum for stimulus frequencies were 1.006 kHz for 1 kHz, 2.012 kHz for 2kHz, and 3.912 kHz for 4 kHz, which means that the oscillation cycle of the mini-drum become short in proportional to the frequency of stimuli. The oscillation cycles of umbo area and the junction area in normal TM for frequencies of the stimuli showed similar integer ratio with the data of latex mini-drum for stimuli less than 4 kHz. In the case of middle ear effusion condition, the Doppler signal showed a tendency of attenuation in all frequencies, which was prominent at 1 kHz and 2 kHz. CONCLUSION: The TM vibration under sound stimulation with frequencies from 1 kHz to 4 kHz in normal and pathologic conditions was demonstrated using signal demodulation method in in vivo condition. The OCT technology could be helpful for functional and structural assessment as an optional modality.

Optical assessment of the in vivo tympanic membrane status using a handheld optical coherence tomography-based otoscope.

OBJECTIVE: Conventional otoscopes and oto-endoscopes, which are used to examine the tympanic membrane (TM), do not provide tomographic information. Optical coherence tomography (OCT) non-invasively reveals the depth-resolved internal microstructure of the TM with very high spatial resolution. We designed this study to examine the TMs with middle ear diseases using a handheld otoscope employing 860 nm spectral domain (SD)-OCT, combined with video camera and to demonstrate the clinical applicability of this system. DESIGN: A total of 120 patients with otologic symptoms were enrolled. TM images were obtained using the handheld OCT-based otoscope (860 nm central wave length, 15 µm axial resolution, 15 µm lateral resolution, and 7 mm scanning range using relay lens). Both OCT and oto-endoscope images were compared according to the clinical characteristics such as perforation, retraction, and postoperative healing process. RESULTS: The objective grade about the thickness of perforation margins and the accurate information about the extent of TM retraction that was not distinguishable by oto-endoscopic exam could be identified using this system. The postoperative healing process of TMs could be also followed using the OCT device. CONCLUSION: These analyses from the surgeon-oriented perspective suggest another useful application of the handheld OCT device.

In vivo 3D imaging of the human tympanic membrane using a wide-field diagonal-scanning optical coherence tomography probe.

A wide-field optical coherence tomography (OCT) probe was developed that adapts a diagonal-scanning scheme for three-dimensional (3D) in vivo imaging of the human tympanic membrane. The probe consists of a relay lens to enhance the lateral scanning range up to 7 mm. Motion artifacts that occur with the use of handheld probes were found to be decreased owing to the diagonal-scanning pattern, which crosses the center of the sample to facilitate entire 3D scans. 3D images could be constructed from a small number of two-dimensional OCT images acquired using the diagonal-scanning technique. To demonstrate the usefulness and performance of the developed system with the handheld probe, in vivo tympanic membranes of humans and animals were imaged in real time.

Treatment of comminuted proximal humeral fractures using locking plate with strut allograft.

BACKGROUND: This study compared the radiologic outcome of fixation using locking plate only with fixation using locking plate with an endosteal strut allograft in the treatment of comminuted proximal humeral fracture. METHODS: Among 52 patients with comminuted proximal humeral fracture, 32 patients underwent fixation with locking plate only, and 20 patients underwent fixation using locking plate with an endosteal strut allograft. The strut allograft was inserted into the intramedullary cavity of the humerus to support the humeral head and fixed with the locking plate. Immediate postoperative radiologic findings were compared with those of 6 months or more after the surgery, and loss of anatomic fixation was defined if the varus malalignment of neck-shaft angle (NSA) was more than 5° or if the change of humeral head height (HHH) was more than 3 mm. RESULTS: In the locking plate-only group, 22 of 32 patients (69%) showed the change in NSA of more than 5°, with an average of 10.2°. The HHH change in 20 patients (62.5%) was more than 3 mm, with an average of 4 mm. Among 20 patients who underwent locking plate with the endosteal strut allograft, the average NSA and HHH change was 3° and 1 mm, respectively. Varus malalignment was evident in 2 patients (10%). The HHH change was more than 3 mm in 1 patient (5%). CONCLUSION: Fixation using a locking plate with an endosteal strut allograft can be considered a reasonable option to maintain the anatomic reduction in elderly patients with comminuted proximal humeral fracture.