A Public BCR Present in a Unique Dual-Receptor-Expressing Lymphocyte from Type 1 Diabetes Patients Encodes a Potent T Cell Autoantigen.

T and B cells are the two known lineages of adaptive immune cells. Here, we describe a previously unknown lymphocyte that is a dual expresser (DE) of TCR and BCR and key lineage markers of both B and T cells. In type 1 diabetes (T1D), DEs are predominated by one clonotype that encodes a potent CD4 T cell autoantigen in its antigen binding site. Molecular dynamics simulations revealed that this peptide has an optimal binding register for diabetogenic HLA-DQ8. In concordance, a synthetic version of the peptide forms stable DQ8 complexes and potently stimulates autoreactive CD4 T cells from T1D patients, but not healthy controls. Moreover, mAbs bearing this clonotype are autoreactive against CD4 T cells and inhibit insulin tetramer binding to CD4 T cells. Thus, compartmentalization of adaptive immune cells into T and B cells is not absolute, and violators of this paradigm are likely key drivers of autoimmune diseases.

A mutagenesis study of autoantigen optimization for potential T1D vaccine design.

A previously reported autoreactive antigen, termed the X-idiotype, isolated from a unique cell population in Type 1 diabetes (T1D) patients, was found to stimulate their CD4+ T cells. This antigen was previously determined to bind more favorably than insulin and its mimic (insulin superagonist) to HLA-DQ8, supporting its strong role in CD4+ T cell activation. In this work, we probed HLA-X-idiotype-TCR binding and designed enhanced-reactive pHLA-TCR antigens using an in silico mutagenesis approach which we functionally validated by cell proliferation assays and flow cytometry. From a combination of single, double, and swap mutations, we identified antigen-binding sites p4 and p6 as potential mutation sites for HLA binding affinity enhancement. Site p6 is revealed to favor smaller but more hydrophobic residues than the native tyrosine, such as valine (Y6V) and isoleucine (Y6I), indicating a steric mechanism in binding affinity improvement. Meanwhile, site p4 methionine mutation to hydrophobic residues isoleucine (M4I) or leucine (M4L) modestly increases HLA binding affinity. Select p6 mutations to cysteine (Y6C) or isoleucine (Y6I) exhibit favorable TCR binding affinities, while a swap p5-p6 tyrosine-valine double mutant (V5Y_Y6V) and a p6-p7 glutamine-glutamine double mutant (Y6Q_Y7Q) exhibit enhanced HLA binding affinity but weakened TCR affinity. This work holds relevance to potential T1D antigen-based vaccine design and optimization.

U-Shape Pillar Strip for 3D Cell-Lumped Organoid Model (3D-COM) Mimicking Lung Cancer Hypoxia Conditions in High-Throughput Screening (HTS).

Hypoxia is a representative tumor characteristic associated with malignant progression in clinical patients. Engineered in vitro models have led to significant advances in cancer research, allowing for the investigation of cells in physiological environments and the study of disease mechanisms and processes with enhanced relevance. In this study, we propose a U-shape pillar strip for a 3D cell-lumped organoid model (3D-COM) to study the effects of hypoxia on lung cancer in a high-throughput manner. We developed a U-pillar strip that facilitates the aggregation of PDCs mixed with an extracellular matrix to make the 3D-COM in 384-plate array form. The response to three hypoxia-activated prodrugs was higher in the 3D-COM than in the 2D culture model. The protein expression of hypoxia-inducible factor 1 alpha (HIF-1α) and HIF-2α, which are markers of hypoxia, was also higher in the 3D-COM than in the 2D culture. The results show that 3D-COM better recapitulated the hypoxic conditions of lung cancer tumors than the 2D culture. Therefore, the U-shape pillar strip for 3D-COM is a good tool to study the effects of hypoxia on lung cancer in a high-throughput manner, which can efficiently develop new drugs targeting hypoxic tumors.

Normal Fermi Surface in the Nodal Superconductor CeCoIn_{5} Revealed via Thermal Conductivity.

The thermal conductivity of heavy-fermion superconductor CeCoIn_{5} was measured with a magnetic field rotating in the tetragonal a-b plane, with the heat current in the antinodal direction, J|| [100]. We observe a sharp resonance in thermal conductivity for the magnetic field at an angle Θ≈12°, measured from the heat current direction [100]. This resonance corresponds to the reported resonance at an angle Θ^{'}≈33° from the direction of the heat current applied along the nodal direction, J||[110]. Both resonances, therefore, occur when the magnetic field is applied in the same crystallographic orientation in the two experiments, regardless of the direction of the heat current, proving conclusively that these resonances are due to the structure of the Fermi surface of CeCoIn_{5}. We argue that the uncondensed Landau quasiparticles, emerging with field, are responsible for the observed resonance. We support our experimental results with density-functional-theory model calculations of the density of states in a rotating magnetic field. Our calculations, using a model Fermi surface of CeCoIn_{5}, reveal several sharp peaks as a function of the field direction. Our study demonstrates that the thermal-conductivity measurement in rotating magnetic field can probe the normal parts of the Fermi surface deep inside the superconducting state.

Ideal chest compression site for cardiopulmonary resuscitation in fontan circulation patients with dextrocardia.

BACKGROUND: We aimed to identify the ideal chest compression site for cardiopulmonary resuscitation (CPR) in patients with a single ventricle with dextrocardia corrected by Fontan surgery. METHODS: The most recent stored chest computed tomography images of all patients with a single ventricle who underwent Fontan surgery were retrospectively analysed. We reported that the ideal chest compression site is the largest part of the compressed single ventricle. To identify the ideal chest compression site, we measured the distance from the midline of the sternum to the point of the maximum sagittal area of the single ventricle as a deviation and calculated the area fraction of the compressed structures. RESULTS: 58 patients (67.2% male) were analysed. The mean right deviation from the midline of the sternum to the ideal compression site was similar to the mean sternum width (32.85 ± 15.61 vs. 31.05 ± 6.75 mm). When chest compression was performed at the ideal site, the area fraction of the single ventricle significantly increased by 7%, which was greater than that of conventional compression (0.15 ± 0.10 vs. 0.22 ± 0.11, P < 0.05). CONCLUSIONS: When performing CPR on a patient with Fontan circulation with dextrocardia, right-sided chest compression may be better than the conventional location.

Comparative Sensing and Judgment Control System for Temperature Maintenance for Optimal Treatment in Hyperthermic Intraperitoneal Chemotherapy Surgery.

For tumors wherein cancer cells remain in the tissue after colorectal cancer surgery, a hyperthermic anticancer agent is injected into the abdominal cavity to necrotize the remaining cancer cells with heat using a hyperthermic intraperitoneal chemotherapy system. However, during circulation, the processing temperature is out of range and the processing result is deteriorated. This paper proposes a look-up table (LUT) module design method that can stably maintain the processing temperature range during circulation via feedback. If the temperature decreases or increases, the LUT transmits a command signal to the heat exchanger to reduce or increase heat input, thereby maintaining the treatment temperature range. The command signal for increasing and decreasing heat input is Tp and Ta, respectively. The command signal for the treatment temperature range is Ts. If drug temperatures below 41 and above 43 °C are input to the LUT, it sends a Tp or Ta signal to the heat exchanger to increase or decrease the input heat, respectively. If the drug's temperature is 41-43 °C, the LUT generates a Ts signal and proceeds with the treatment. The proposed system can automatically control drug temperature using temperature feedback to ensure rapid, accurate, and safe treatment.

Adjustment of the Length Variation With Wire-driven Using the Stand Looper Tension Technique for Surgical Robot Applications.

MOTIVATION: The wire-driven method used in the field of surgical robots has the advantage of light weight. However, in the process of pull and push for the operation of forceps, the length of the wire is not match, causing malfunction. To solve this problem, the application of looper-tension technology would be suitable. This paper contributes to adjusting the length of the wire by inserting a stand between the wire-driven joints and adding a looper-tension between the stands to adjust the rotation radius of the roll. METHODS: The method consisting of three rolls and loopers for connection between the stands minimizes errors by adjusting the length of the loop in a balanced state due to the rotation change of the roll during the pull and push of the robot arm. The angle and tension applied to the looper are 25° and 8.6 MPa, respectively. RESULTS: An output response can be obtained when the reference operating point fluctuates by ± 50% of the input angle and tension, and if the reference operating point fluctuates by ± 30% while the input angle and tension are fixed, the output response occurs oppositely. When a .15 kg object is loaded up/down with 1.5 newton using forceps, the change in length of pull and push coincides. CONCLUSION: The advantage is that the error of wire pull, and push operation can be reduced, and accurate operation can be expected. Since the proposed technology is applied between joints, the integration process is not complicated, and the weight is light.

A Method to Reduce Tension Differences in Pull/Push Manipulation for a Robot in Fluorescence Emission-Guided Surgical Microscopy.

MOTIVATION: A fluorescence emission-guided microscope used to monitor the outcome of cancer removal surgery is highly effective when employing a manipulator to motorize and switch the observation direction. It is necessary to minimize the alignment of looper tension between the stands for pull/push to change the direction of the manipulator and reduce the error rate caused by tension differences. This paper presents a method to minimize the error rate of looper tension between the stands. METHODS: \The looper is inserted between the stands of the manipulator to minimize the difference in tension and make the stress on the pull and push of the looper constant. The constant stress allows the manipulator to move stably in left/right, up/down, and left/right movements, which will be effective for full-camera observation and close-up shots of the end effector. RESULTS: Reducing the tolerance for differences in the manipulator's looper tension (angle and tension) is crucial. When the input value of the looper tension angle is 50°, the output should closely match 50°. Consequently, the measured response has a tolerance of ±49.98%, resulting in an error rate of .02% (1/50th level). CONCLUSION: A method is proposed to minimize the error rate of the manipulator's looper tension in a robot-based fluorescence emission-guided microscope used to observe the status of cancer surgery. As a result, a stable manipulator with a minimal error rate can achieve a 3.986x magnification for close-up observation by switching between high and low orientations.

Noncanonical mode of ERK action controls alternative αß and γδ T cell lineage fates.

Gradations in extracellular regulated kinase (ERK) signaling have been implicated in essentially every developmental checkpoint or differentiation process encountered by lymphocytes. Yet, despite intensive effort, the molecular basis by which differences in ERK activation specify alternative cell fates remains poorly understood. We report here that differential ERK signaling controls lymphoid-fate specification through an alternative mode of action. While ERK phosphorylates most substrates, such as RSK, by targeting them through its D-domain, this well-studied mode of ERK action was dispensable for development of γδ T cells. Instead, development of γδ T cells was dependent upon an alternative mode of action mediated by the DEF-binding pocket (DBP) of ERK. This domain enabled ERK to bind a distinct and select set of proteins required for specification of the γδ fate. These data provide the first in vivo demonstration for the role of DBP-mediated interactions in orchestrating alternate ERK-dependent developmental outcomes.

The Design of an Automatic Temperature Compensation System through Smart Heat Comparison/Judgment and Control for Stable Thermal Treatment in Hyperthermic Intraperitoneal Chemotherapy (HIPEC) Surgery.

After surgery for ovarian cancer or colorectal cancer, residual tumors are left around. A practical way to treat residual tumors is to destroy them with heat by injecting high-temperature drugs into the abdominal cavity. The injected medicinal substances are induced to flow out of the abdominal cavity; then, the spilled drug flows back into the abdominal cavity through feedback. During this process, the heat starts to decrease; thus, the treatment performance reduces. To overcome this problem, this study compares and assesses the temperature needed to maintain the heat for treatment and transmits a command signal to the heat exchanger through a look-up table (LUT). When the temperature decreases during the circulation of medications leaking out of the abdominal cavity, the LUT transmits a control signal (Tp) to the heat exchanger, which increases or vice versa. However, if the temperature (To) is within the treatment range, the LUT sends a Ts signal to the heat exchanger. This principle generates a pulse signal for the temperature difference (Tdif) in TC by comparing and determining the temperature (To) of the substance flowing out of the abdominal cavity with the reference temperature (Tref) through the temperature comparator (TC). At this time, if the signal is 41 °C or less, the LUT generates (heats) a Tp signal so that the temperature of the heat exchanger can be maintained in the range of 41 °C to 43 °C. If the Tdif is 44 °C or higher, the LUT generates (cools) the Ta signal and maintains the temperature of the heat exchanger at 41-43 °C. If the Tdif is maintained at 41-43 °C, the LUT generates a Tx signal to stop the system performance. At this time, the TC operation performance and Tdif generation process for comparing and determining the signal of To and Tref for drugs leaking out of the abdominal cavity is very important. It was observed that the faster the response signal, the lower the comparison and judgment error was; therefore, the response signal was confirmed to be 0.209 µs. The proposed method can guarantee rapid/accurate/safe treatment and automatically induce temperature adjustment; thus, it could be applied to the field of surgery.

Design of a Compact Hologram System Capable of 3D Lesion Diagnosis in Clinic.

MOTIVATION: This paper proposes a small-sized hologram system for the 3D imaging of lesions in a clinical environment. In a general hologram system, the distance between the beam-generating device and the screen (400 mm) and the size of the screen must be increased proportionally to obtain excellent image quality. However, in a clinical environment, the beam spread distance and screen size must be reduced. This paper proposes a method for reducing the beam divergence distance and screen size for clinical applications. METHODS: To reduce the beam spread distance and screen size, a beam prism with a 45° refractive index is used to reduce the beam spread distance by 1/3. The direction of the bent light must be adjusted such that it can reach the screen accurately. However, because the reflected light may be refracted owing to the material properties of the mirror and cause loss, this problem can be solved by using a full reflection mirror. RESULTS: The beam spread distance of the designed hologram system is 200 mm. The types of lesions obtained from the 3D images of the hologram include the lung, liver, and colon. The image resolution is 300 × 145. CONCLUSION: If the proposed method is used in a clinical environment, doctors can improve their understanding of the patient quickly and efficiently; thereby, shortening the treatment time. The proposed hologram system is expected to be useful in treatment rooms, operating rooms, and educational programs in medical schools.

Design of a Miniature Observation Robot for Light Emitting Diode Irradiation and Indocyanine Green Fluorescence-emission Guided Lymph Node Monitoring in Operating Rooms.

MOTIVATION: Typical surgical microscopes used for fluorescence-based lymph node detection experience limitations such as weight and restricted adjustability of the integrated light emitting diode (LED) and camera. This restricts the capture of detailed images of specific regions within the lesion. RESEARCH GOAL: This study proposes a miniature observation robot design that offers adjustable working distance (WD) and rotational radius, along with zoom-in/zoom-out functionality. METHODS: A five-degree-of-freedom manipulator was designed, with the end effector incorporating an LED and concave lens to widen the beam width for comprehensive lesion illumination. Additionally, a long-pass filter was integrated into the camera system to enhance image resolution. EXPERIMENTAL RESULTS: Experiments were conducted using a fluorescence-expressing phantom to evaluate the performance of the robot. Results demonstrated a captured image resolution of 9600 × 3240 pixels and a zoom-in/zoom-out capacity of up to 3.68 times. CONCLUSION: The proposed robot design is cost-effective and highly adjustable, enabling suitability for rapid and accurate detection of fresh lymph nodes during surgeries. The robot's capability to detect small lesions (<1 cm), as validated by phantom tests, holds promise for the detection of minute lymph nodes.

Cell Proliferation Receptor-Enhanced 3D High-Throughput Screening Model for Optimized Drug Efficacy Evaluation in Breast Cancer Cells.

A higher correlation of epidermal growth factor receptor (EGFR)-targeting drugs has been reported with the use of the cell proliferation receptor-enhanced three-dimensional high-throughput screening model (CPRE 3D-HTS model) compared with two-dimensional (2D) cell-based HTS. A greater expression of differential human EGFR 2 (HER2) protein between HER2-positive and HER2-negative cell lines was observed in breast cancer (BC) cell lines cultured using the CPRE 3D-HTS model compared with 2D-cultured cells. When using 2D-cultured cells, properties such as the expression of the cell proliferation receptor are lost as the cells attach to the bottom of the well plate. In an effort to solve this problem, the CPRE 3D-HTS model expressing high cell proliferation receptors was optimized by the selection of alginate as the extracellular matrix. Results from the use of the CPRE 3D-HTS model showed higher drug resistance with increased expression of drug resistance-related proteins. Of particular interest, a higher correlation of HER2-targeted drugs was observed with the use of the CPRE 3D-HTS model. In order to validate this higher correlation of target drugs observed in the CPRE 3D-HTS model, the results of Western blot analysis and high content imaging analysis were analyzed, which confirmed that 3D-cultured BC cell lines showed a greater difference in the expression of HER2-positive and HER2-negative BC cell lines than 2D-cultured cells. Thus, the use of CPRE 3D-HTS using a 384-pillar plate resulted in increased accuracy when screening HER2-targeted drugs in BC, and it is a very useful platform for analyzing the efficacy of targeted drugs by enhancing the expression of HER2.

Simplified in vitro 3D co-culture-based blood-brain barrier model using transwell.

The blood-brain barrier (BBB) is a major hurdle for treatment of brain diseases. To overcome this, precise and reproducible BBB model is one of the key factors for successful evaluation of BBB-penetrating efficacy of developmental drugs. Thus, in vitro BBB model recapitulating the physiological structure of the BBB is a valuable tool for drug discovery and development for brain diseases. Here, we develop a simplified 3D co-culture-based BBB model using immortalized human brain endothelial cells and immortalized human astrocytes mixed with Matrigel allowing model preparation within 30 min. We directly compare our 3D BBB model to a 2D BBB model comprised solely of immortalized brain endothelial cells, to demonstrate that our 3D BBB model blocks penetration of Dextran molecules with various molecular weights, remain durable and impermeable even in a BBB-degrading condition, and rapidly form tight junctions while the 2D BBB model do not. In conclusion, this establishes our simplified 3D BBB model as a valuable tool for high throughput screening of drug candidates for brain diseases.

Functional validation of the simplified in vitro 3D Co-culture based BBB model.

Various methods of generating 2D and 3D in vitro blood-brain barrier (BBB) models have previously been published with the objective of developing therapeutics for brain diseases. In general, published methods including our published method demonstrate that in vivo-like semi-permeable barrier can be generated. To further verify that an in vitro BBB model closely represents BBB, functional validation is required. Here, we functionally validate our in vitro 3D BBB model using rituximab as a representative therapeutic antibody and previously published anti-TfR (transferrin receptor) antibodies as representative BBB-penetrating antibodies. We demonstrate that our BBB model can efficiently block rituximab while allowing receptor-mediated transcytosis (RMT) of anti-TfR antibodies. In addition, we showed that RMT efficacy of anti-TfR antibodies with different binding affinity can be displayed using our BBB model. In conclusion, this demonstrates that our BBB model functionally mimics the BBB as well as having BBB-like physical properties, further establishing our BBB model as a screening tool for discovery and development of therapeutics for brain diseases.

Speed Limit for a Highly Irreversible Process and Tight Finite-Time Landauer's Bound.

Landauer's bound is the minimum thermodynamic cost for erasing one bit of information. As this bound is achievable only for quasistatic processes, finite-time operation incurs additional energetic costs. We find a tight finite-time Landauer's bound by establishing a general form of the classical speed limit. This tight bound well captures the divergent behavior associated with the additional cost of a highly irreversible process, which scales differently from a nearly irreversible process. We also find an optimal dynamics which saturates the equality of the bound. We demonstrate the validity of this bound via discrete one-bit and coarse-grained bit systems. Our Letter implies that more heat dissipation than expected occurs during high-speed irreversible computation.

Specular Reflection Suppression through the Adjustment of Linear Polarization for Tumor Diagnosis Using Fluorescein Sodium.

In tumor surgery, the edges of the tumor can be visually observed using a fluorescent contrast agent and a fluorescent imaging device. By distinguishing it from normal tissues and blood vessels, it is possible to objectively judge the extent of resection while visually observing it during surgery, and it guarantees safe tumor resection based on more information. However, the main problem of such an imaging device is the specular reflection phenomenon. If specular reflection overlaps with important lesion locations, they are a major factor leading to diagnostic errors. Here, we propose a method to reduce specular reflection that occurs during tumor diagnosis using a linear polarization filter and fluorescent contrast agent. To confirm the effect of removing specular reflection, a self-made fluorescein sodium vial phantom was used, and the reliability of the results was increased using a large animal (pig) test. As a result of the experiment, it was possible to obtain an image in which specular reflection was removed by controlling the rotation angle of the filter by 90° and 270°, and the same results were confirmed in the phantom experiment and the animal experiment.

Strong Zero-Phonon Transition from Point Defect-Stacking Fault Complexes in Silicon Carbide Nanowires.

Crystallographic defects such as vacancies and stacking faults engineer electronic band structure at the atomic level and create zero- and two-dimensional quantum structures in crystals. The combination of these point and planar defects can generate a new type of defect complex system. Here, we investigate silicon carbide nanowires that host point defects near stacking faults. These point-planar defect complexes in the nanowire exhibit outstanding optical properties of high-brightness single photons (>360 kcounts/s), a fast recombination time (<1 ns), and a high Debye-Waller factor (>50%). These distinct optical properties of coupled point-planar defects lead to an unusually strong zero-phonon transition, essential for achieving highly efficient quantum interactions between multiple qubits. Our findings can be extended to other defects in various materials and therefore offer a new perspective for engineering defect qubits.

High Throughput 3D Cell Migration Assay Using Micropillar/Microwell Chips.

The 3D cell migration assay was developed for the evaluation of drugs that inhibit cell migration using high throughput methods. Wound-healing assays have commonly been used for cell migration assays. However, these assays have limitations in mimicking the in vivo microenvironment of the tumor and measuring cell viability for evaluation of cell migration inhibition without cell toxicity. As an attempt to manage these limitations, cells were encapsulated with Matrigel on the surface of the pillar, and an analysis of the morphology of cells attached to the pillar through Matrigel was performed for the measurement of cell migration. The micropillar/microwell chips contained 532 pillars and wells, which measure the migration and viability of cells by analyzing the roundness and size of the cells, respectively. Cells seeded in Matrigel have a spherical form. Over time, cells migrate through the Matrigel and attach to the surface of the pillar. Cells that have migrated and adhered have a diffused shape that is different from the initial spherical shape. Based on our analysis of the roundness of the cells, we were able to distinguish between the diffuse and spherical shapes. Cells in Matrigel on the pillar that were treated with migration-inhibiting drugs did not move to the surface of the pillar and remained in spherical forms. During the conduct of experiments, 70 drugs were tested in single chips and migration-inhibiting drugs without cell toxicity were identified. Conventional migration assays were performed using transwell for verification of the four main migration-inhibiting drugs found on the chip.

Mid-term outcomes of the Pulsta transcatheter pulmonary valve for the native right ventricular outflow tract.

OBJECTIVES: The aim of this study is to present the mid-term outcomes of Pulsta valve. BACKGROUND: The Pulsta valve is a Self-expandable knitted nitinol-wire stent mounted with a treated tri-leaflet α-Gal-free porcine pericardial valve for percutaneous pulmonary valve implantation (PPVI) in patients with native right ventricular outflow tract (RVOT) lesions. METHODS: A multi-center clinical trial using Pulsta valve® was designed for patients with severe pulmonary regurgitation (PR) in the native RVOT in multiple centers in South Korea and 25 patients were enrolled. Before PPVI, severe PR (mean PR fraction: 45.5 ± 6.9%) and enlarged RV volume (mean indexed RV end-diastolic volume; 169.7 ± 13.0 ml/m2 ) was present. The mean age was 21.6 ± 6.6 years old. RESULTS: All patients were successfully implanted with 26, 28, or 32 mm diameter of Pulsta valve loaded on the 18 or 20 French delivery catheters. At 6 months follow up, indexed RV end-diastolic volume was decreased to 126.9 ± 16.9 ml/m2 . At mean 33.1 ± 14.3 months follow-up, the mean value of mean pressure gradient in Pulsta valve was 6.5 ± 3.0 mmhg without significant PR. There was no serious device-related adverse event. CONCLUSIONS: A multi-center clinical trial was completed successfully with planned Pulsta valve implantation and demonstrated good mid-term effectiveness without device-related serious adverse events.