The intercentriolar fibers function as docking sites of centriolar satellites for cilia assembly.

Two mother centrioles in an animal cell are linked by intercentriolar fibers that have CROCC/rootletin as their main building block. Here, we investigated the regulatory role of intercentriolar/rootlet fibers in cilia assembly. The cilia formation rates were significantly reduced in the CEP250/C-NAP1 and CROCC/rootletin knockout (KO) cells, irrespective of the departure of the young mother centrioles from the basal bodies. In addition, centriolar satellites were dispersed throughout the cytoplasm in the CEP250 and CROCC KO cells. We observed that PCM1 directly binds to CROCC. Their interaction is critical not only for the accumulation of centriolar satellites near the centrosomes/basal bodies but also for cilia formation. Finally, we observed that the centriolar satellite proteins are localized at the intercentriolar/rootlet fibers in the kidney epithelial cells. Based on these findings, we propose that the intercentriolar/rootlet fibers function as docking sites for centriolar satellites near the centrosomes/basal bodies and facilitate the cilia assembly process.

Assessment of Image Quality of Coronary CT Angiography Using Deep Learning-Based CT Reconstruction: Phantom and Patient Studies.

BACKGROUND: In coronary computed tomography angiography (CCTA), the main issue of image quality is noise in obese patients, blooming artifacts due to calcium and stents, high-risk coronary plaques, and radiation exposure to patients. OBJECTIVE: To compare the CCTA image quality of deep learning-based reconstruction (DLR) with that of filtered back projection (FBP) and iterative reconstruction (IR). METHODS: This was a phantom study of 90 patients who underwent CCTA. CCTA images were acquired using FBP, IR, and DLR. In the phantom study, the aortic root and the left main coronary artery in the chest phantom were simulated using a needleless syringe. The patients were classified into three groups according to their body mass index. Noise, the signal-to-noise ratio (SNR), and the contrast-to-noise ratio (CNR) were measured for image quantification. A subjective analysis was also performed for FBP, IR, and DLR. RESULTS: According to the phantom study, DLR reduced noise by 59.8% compared to FBP and increased SNR and CNR by 121.4% and 123.6%, respectively. In a patient study, DLR reduced noise compared to FBP and IR. Furthermore, DLR increased the SNR and CNR more than FBP and IR. In terms of subjective scores, DLR was higher than FBP and IR. CONCLUSION: In both phantom and patient studies, DLR effectively reduced image noise and improved SNR and CNR. Therefore, the DLR may be useful for CCTA examinations.

NSD Overexpression in the Fat Body Increases Antimicrobial Peptide Production by the Immune Deficiency Pathway in Drosophila.

Nuclear receptor-binding SET domain-containing protein 1 (NSD1) inactivation in tumor cells contributes to an immune-cold phenotype, indicating its potential association with immune disturbances. Drosophila NSD is a homolog of the human NSD1. Thus, in this study, we investigated the effect of NSD overexpression in the fat body, the central organ involved in Drosophila immune responses. Upon ectopic expression of NSD in the fat body, the mRNA levels of antimicrobial peptides increased. Using reporter constructs containing deletions of various NF-κB sites in the Attacin-A (AttA) promoter, we found that transcriptional activation by NSD is mainly mediated via the IMD pathway by activating Relish. Since the IMD pathway is required to resist Gram-negative bacterial infections, we further examined the effect of fat body-specific NSD overexpression on Drosophila immune defenses. Upon oral ingestion of Gram-negative Pseudomonas entomophila, the survival rate of the NSD-overexpressing larvae was higher than that of the wild type, suggesting a positive role of NSD in immune responses. Taken together, these results suggest the association of NSD with the IMD pathway and is thus expected to contribute to the elucidation of the molecular mechanisms of immune malfunction in various NSD1-associated human diseases.

Identification of a Structurally Dynamic Domain for Oligomer Formation in Rootletin.

Rootletin is the main component of the ciliary rootlet and functions as a centriole linker connecting the two mother centrioles. Despite the functional importance of rootletin, the molecular architecture of the rootletin filament and its assembly mechanism are poorly understood. Here, we identify the coiled-coil domain 3 (CCD3) of rootletin as the key domain for its cellular function. The crystal structure of the CCD31108-1317 fragment containing 28 heptad repeats and 1 hendecad repeat reveals that it forms a parallel coiled-coil dimer spanning approximately 300 Šin length. Crosslinking experiments and biophysical analyses of the minimal functional region of CCD3 (CCD3-6) suggest that CCD3-6 is structurally dynamic and may be important for oligomer formation. We also show that oligomerization-defective CCD3 mutants fail in centrosomal localization and centriole linkage, suggesting that rootletin oligomerization may be important for its function.