Phase Separation and Protein Partitioning in Compartmentalized Cell-Free Expression Reactions.

Liquid-liquid phase separation (LLPS) is important to control a wide range of reactions from gene expression to protein degradation in a cell-sized space. To bring a better understanding of the compatibility of such phase-separated structures with protein synthesis, we study emergent LLPS in a cell-free transcription-translation (TXTL) reaction. When the TXTL reaction composed of many proteins is concentrated, the uniformly mixed state becomes unstable, and membrane-less phases form spontaneously. This LLPS droplet formation is induced when the TXTL reaction is enclosed in water-in-oil emulsion droplets, in which water evaporates from the surface. As the emulsion droplets shrink, smaller LLPS droplets appear inside the emulsion droplets and coalesce into large phase-separated domains that partition the localization of synthesized reporter proteins. The presence of PEG in the TXTL reaction is important not only for versatile cell-free protein synthesis but also for the formation of two large domains capable of protein partitioning. Our results may shed light on the dynamic interplay of LLPS formation and cell-free protein synthesis toward the construction of synthetic organelles.

Comparison of three artificial intelligence algorithms for automatic cobb angle measurement using teaching data specific to three disease groups.

Spinal deformities, including adolescent idiopathic scoliosis (AIS) and adult spinal deformity (ASD), affect many patients. The measurement of the Cobb angle on coronal radiographs is essential for their diagnosis and treatment planning. To enhance the precision of Cobb angle measurements for both AIS and ASD, we developed three distinct artificial intelligence (AI) algorithms: AIS/ASD-trained AI (trained with both AIS and ASD cases); AIS-trained AI (trained solely on AIS cases); ASD-trained AI (trained solely on ASD cases). We used 1612 whole-spine radiographs, including 1029 AIS and 583 ASD cases with variable postures, as teaching data. We measured the major and two minor curves. To assess the accuracy, we used 285 radiographs (159 AIS and 126 ASD) as a test set and calculated the mean absolute error (MAE) and intraclass correlation coefficient (ICC) between each AI algorithm and the average of manual measurements by four spine experts. The AIS/ASD-trained AI showed the highest accuracy among the three AI algorithms. This result suggested that learning across multiple diseases rather than disease-specific training may be an efficient AI learning method. The presented AI algorithm has the potential to reduce errors in Cobb angle measurements and improve the quality of clinical practice.

Comparative Study of S2-Alar-Iliac Screw Trajectories between Males and Females Using Three-Dimensional Computed Tomography Analysis: The True Lateral Angulation of the S2-Alar-Iliac Screw in the Axial Plane.

The S2 alar-iliac screw (S2AIS) is commonly used for long spinal fusion as a rigid distal foundation in spinal deformity surgeries, and it is also used in percutaneous sacropelvic fixation for providing an in-line connection to the proximal spinal constructs without using offset connectors. Although the pelvic shape is different between males and females, reports on S2AIS trajectories according to gender have been scarce in the literature. In this paper, S2AIS trajectories are compared between males and females using pelvic three-dimensional computed tomography (3D-CT) in a normal Japanese population. After resetting the caudal angulation in CT-imaging plane manipulation, the angulation of S2AIS was more lateral in the axial plane and more horizontal in the coronal plane in females. Mean distances from the midline to starting points of S2AIS tended to be shorter in females, whereas mean distances from the midline to the posterior superior iliac spine was significantly longer in females. We also found that there were positive correlations between the patients' height and the maximal lengths of S2AISs, and the patients' height and minimal areas of S2AIS pathways. Our results are useful not only for conventional open spinal surgery, but also for minimally invasive spine surgery.

Posterior and Anterior Fusion for Severe Cervical Kyphosis in a Patient with Chondrodysplasia Punctata: A Case Report.

CASE: A 4-year-old girl with spastic gait and hand clumsiness who was diagnosed with cervical myelopathy caused by atlantoaxial dislocation and midcervical severe kyphosis associated with chondrodysplasia punctata (CDP). The patient underwent posterior instrumentation and anterior spinal fusion and successful correction with osseous fusion was obtained 8 months after surgery. In addition, the preoperative neurological symptoms were completely recovered. CONCLUSION: Owing to the characteristics of CDP, the treatment for the cervical lesion is extremely complicated. Successful stabilization and improvement of the neurological symptom were achieved by combining posterior and anterior fusion with instrumentation in this case.