Endosulfine alpha maintains spindle pole integrity by recruiting Aurora A during mitosis.

BACKGROUND: The maintenance of spindle pole integrity is essential for spindle assembly and chromosome segregation during mitosis. However, the underlying mechanisms governing spindle pole integrity remain unclear. METHODS: ENSA was inhibited by siRNA or MKI-2 treatment and its effect on cell cycle progression, chromosome alignment and microtubule alignment was observed by immunohistochemical staining and western blotting. PP2A-B55α knockdown by siRNA was performed to rescue the phenotype caused by ENSA inhibition. The interaction between ENSA and Aurora A was detected by in situ PLA. Furthermore, orthotopic implantation of 4Tl-luc cancer cells was conducted to confirm the consistency between the in vitro and in vivo relationship of the ENSA-Aurora A interaction. RESULTS: During mitosis, p-ENSA is localized at the spindle poles, and the inhibition of ENSA results in mitotic defects, such as misaligned chromosomes, multipolar spindles, asymmetric bipolar spindles, and centrosome defects, with a delay in mitotic progression. Although the mitotic delay caused by ENSA inhibition was rescued by PP2A-B55α depletion, spindle pole defects persisted. Notably, we observed a interaction between ENSA and Aurora A during mitosis, and inhibition of ENSA reduced Aurora A expression at the mitotic spindle poles. Injecting MKI-2-sensitized tumors led to increased chromosomal instability and downregulation of the MASTL-ENSA-Aurora A pathway in an orthotopic breast cancer mouse model. CONCLUSIONS: These findings provide novel insights into the regulation of spindle pole integrity by the MASTL-ENSA-Aurora A pathway during mitosis, highlighting the significance of ENSA in recruiting Aurora A to the spindle pole, independent of PP2A-B55α.

Evaluation of artificial intelligence model for crowding categorization and extraction diagnosis using intraoral photographs.

Determining the severity of dental crowding and the necessity of tooth extraction for orthodontic treatment planning are time-consuming processes and there are no firm criteria. Thus, automated assistance would be useful to clinicians. This study aimed to construct and evaluate artificial intelligence (AI) systems to assist with such treatment planning. A total of 3,136 orthodontic occlusal photographs with annotations by two orthodontists were obtained. Four convolutional neural network (CNN) models, namely ResNet50, ResNet101, VGG16, and VGG19, were adopted for the AI process. Using the intraoral photographs as input, the crowding group and the necessity of tooth extraction were obtained. Arch length discrepancy analysis with AI-detected landmarks was used for crowding categorization. Various statistical and visual analyses were conducted to evaluate the performance. The maxillary and mandibular VGG19 models showed minimum mean errors of 0.84 mm and 1.06 mm for teeth landmark detection, respectively. Analysis of Cohen's weighted kappa coefficient indicated that crowding categorization performance was best in VGG19 (0.73), decreasing in the order of VGG16, ResNet101, and ResNet50. For tooth extraction, the maxillary VGG19 model showed the highest accuracy (0.922) and AUC (0.961). By utilizing deep learning with orthodontic photographs, dental crowding categorization and diagnosis of orthodontic extraction were successfully determined. This suggests that AI can assist clinicians in the diagnosis and decision making of treatment plans.

Feasibility of artificial intelligence-based decision supporting system in tolvaptan prescription for autosomal dominant polycystic kidney disease.

PURPOSE: Total kidney volume (TKV) measurement is crucial for selecting treatment candidates in autosomal dominant polycystic kidney disease (ADPKD). We developed and investigated the performance of fully-automated 3D-volumetry model and applied it to software as a service (SaaS) for clinical support on tolvaptan prescription in ADPKD patients. MATERIALS AND METHODS: Computed tomography scans of ADPKD patients taken between January 2000 and June 2022 were acquired from seven institutions. The quality of the images was manually reviewed in advance. The acquired dataset was split into training, validation, and test datasets at a ratio of 8.5:1:0.5. Convolutional, neural network-based automatic segmentation model was trained to obtain 3D segment mask for TKV measurement. The algorithm consisted of three steps: data preprocessing, ADPKD area extraction, and post-processing. After performance validation with the Dice score, 3D-volumetry model was applied to SaaS which is based on Mayo imaging classification for ADPKD. RESULTS: A total of 753 cases with 95,117 slices were included. The differences between the ground-truth ADPKD kidney mask and the predicted ADPKD kidney mask were negligible, with intersection over union >0.95. The post-process filter successfully removed false alarms. The test-set performance was homogeneously equal and the Dice score of the model was 0.971; after post-processing, it improved to 0.979. The SaaS calculated TKV from uploaded Digital Imaging and Communications in Medicine images and classified patients according to height-adjusted TKV for age. CONCLUSIONS: Our artificial intelligence-3D volumetry model exhibited effective, feasible, and non-inferior performance compared with that of human experts and successfully predicted the rapid ADPKD progressor.

Three-dimensional dental model constructed from an average dental form.

INTRODUCTION: The purposes of this study were (1) to discover a simple method for computing an average dental form, (2) to evaluate the validity of the new computing method, and (3) to build an average digital dental model based on the resulting average dental form. METHODS: Dental casts of 51 adults were scanned, and 3-dimensional virtual models were reconstructed by using specialized software. Fifty-two anatomic landmarks were plotted on each model to obtain a configuration representing that model. The averaging method was a simplified approach of the generalized partial Procrustes analysis (GPPA); called multiple partial Procrustes analysis (MPPA), it was used to superimpose the 51 sample configurations. For the evaluation method, the GPPA was performed to estimate the population mean form. This mean form was compared with the average forms obtained from the proposed method by using the Procrustes distance as the measure of difference. Construction of the average dental model was based on an average form obtained from the proposed method, and a 3-dimensional dental mesh model was reconstructed. RESULTS: The average forms from the proposed method showed no significant differences compared with the one from the GPPA. The Procrustes distances were about 0.01 mm, which can safely be considered insignificant. CONCLUSIONS: The proposed averaging method, which is simpler than the traditional GPPA approach, was found to be feasible for computing average dental forms. The average dental model constructed in this study might serve as a guide for treatment planning in orthodontic and prosthodontic dentistry.

Auditory Behavior in Adult-Blinded Mice.

Cross-modal plasticity occurs when the function of remaining senses is enhanced following deprivation or loss of a sensory modality. Auditory neural responses are enhanced in the auditory cortex, including increased sensitivity and frequency selectivity, following short-term visual deprivation in adult mice (Petrus et al. Neuron 81:664-673, 2014). Whether or not these visual deprivation-induced neural changes translate into improved auditory perception and performance remains unclear. As an initial investigation of the effects of adult visual deprivation on auditory behaviors, CBA/CaJ mice underwent binocular enucleation at 3-4 weeks old and were tested on a battery of learned behavioral tasks, acoustic startle response (ASR), and prepulse inhibition (PPI) tests beginning at least 2 weeks after the enucleation procedure. Auditory brain stem responses (ABRs) were also measured to screen for potential effects of visual deprivation on non-behavioral hearing function. Control and enucleated mice showed similar tone detection sensitivity and frequency discrimination in a conditioned lick suppression test. Both groups showed normal reactivity to sound as measured by ASR in a quiet background. However, when startle-eliciting stimuli were presented in noise, enucleated mice showed decreased ASR amplitude relative to controls. Control and enucleated mice displayed no significant differences in ASR habituation, PPI tests, or ABR thresholds, or wave morphology. Our findings suggest that while adult-onset visual deprivation induces cross-modal plasticity at the synaptic and circuit levels, it does not substantially influence simple auditory behavioral performance.

Characterization and Determination of Nanoparticles in Commercial Processed Foods.

A wide variety of foods manufactured by nanotechnology are commercially available on the market and labeled as nanoproducts. However, it is challenging to determine the presence of nanoparticles (NPs) in complex food matrices and processed foods. In this study, top-down-approach-produced (TD)-NP products and nanobubble waters (NBWs) were chosen as representative powdered and liquid nanoproducts, respectively. The characterization and determination of NPs in TD-NP products and NBWs were carried out by measuring constituent particle sizes, hydrodynamic diameters, zeta potentials, and surface chemistry. The results show that most NBWs had different characteristics compared with those of conventional sparkling waters, but nanobubbles were unstable during storage. On the other hand, powdered TD-NP products were found to be highly aggregated, and the constituent particle sizes less than 100 nm were remarkably observed after dispersion compared with counterpart conventional bulk-sized products by scanning electron microscopy at low acceleration voltage and cryogenic transmission electron microscopy. The differences in chemical composition and chemical state between TD-NPs and their counterpart conventional bulk products were also found by X-ray photoelectron spectroscopy. These findings will provide basic information about the presence of NPs in nano-labeled products and be useful to understand and predict the potential toxicity of NPs applied to the food industry.

Improving the Stability and Curcumin Retention Rate of Curcumin-Loaded Filled Hydrogel Prepared Using 4αGTase-Treated Rice Starch.

In this study, 4-α-glucanotransferase (4αGTase)-treated rice starch (GS) was added after 1-h (1 GS) and 96-h (96 GS) treatments to the aqueous phase of a curcumin-loaded emulsion to produce filled hydrogels (1 GS-FH and 96 GS-FH, respectively). The relative protective effects of the FH system, native rice starch-based filled hydrogel (RS-FH), and emulsion without starch (EM), on curcumin were evaluated based on ultraviolet (UV) stability and simulated gastrointestinal studies. The UV stability and curcumin retention after in vitro digestion of the filled hydrogels (FH) samples were greater than those of the EM samples. RS-FH showed a 2.28-fold improvement in UV stability over EM due to the higher viscosity of RS. 1 GS-FH and 96 GS-FH increased curcumin retention by 2.31- and 2.60-fold, respectively, and the microstructure of 96 GS-FH, determined using confocal laser microscopy, remained stable even after the stomach phase. These effects were attributed to the molecular structure of GS, with decreased amylopectin size and amylose content resulting from the enzyme treatment. The encapsulation of lipids within the GS hydrogel particles served to protect and deliver the curcumin component, suggesting that GS-FH can be applied to gel-type food products and improve the chemical stability of curcumin.

Influence of the Depth of the Convolutional Neural Networks on an Artificial Intelligence Model for Diagnosis of Orthognathic Surgery.

The aim of this study was to investigate the relationship between image patterns in cephalometric radiographs and the diagnosis of orthognathic surgery and propose a method to improve the accuracy of predictive models according to the depth of the neural networks. The study included 640 and 320 patients requiring non-surgical and surgical orthodontic treatments, respectively. The data of 150 patients were exclusively classified as a test set. The data of the remaining 810 patients were split into five groups and a five-fold cross-validation was performed. The convolutional neural network models used were ResNet-18, 34, 50, and 101. The number in the model name represents the difference in the depth of the blocks that constitute the model. The accuracy, sensitivity, and specificity of each model were estimated and compared. The average success rate in the test set for the ResNet-18, 34, 50, and 101 was 93.80%, 93.60%, 91.13%, and 91.33%, respectively. In screening, ResNet-18 had the best performance with an area under the curve of 0.979, followed by ResNets-34, 50, and 101 at 0.974, 0.945, and 0.944, respectively. This study suggests the required characteristics of the structure of an artificial intelligence model for decision-making based on medical images.

Discovery and Characterization of a Novel MASTL Inhibitor MKI-2 Targeting MASTL-PP2A in Breast Cancer Cells and Oocytes.

Although microtubule-associated serine/threonine kinase-like (MASTL) is a promising target for selective anticancer treatment, MASTL inhibitors with nano range potency and antitumor efficacy have not been reported. Here, we report a novel potent and selective MASTL inhibitor MASTL kinase inhibitor-2 (MKI-2) identified in silico through a drug discovery program. Our data showed that MKI-2 inhibited recombinant MASTL activity and cellular MASTL activity with IC50 values of 37.44 nM and 142.7 nM, respectively, in breast cancer cells. In addition, MKI-2 inhibited MASTL kinase rather than other AGC kinases, such as ROCK1, AKT1, PKACα, and p70S6K. Furthermore, MKI-2 exerted various antitumor activities by inducing mitotic catastrophe resulting from the modulation of the MASTL-PP2A axis in breast cancer cells. The MKI-2 treatment showed phenocopies with MASTL-null oocyte in mouse oocytes, which were used as a model to validate MKI-2 activity. Therefore, our study provided a new potent and selective MASTL inhibitor MKI-2 targeting the oncogenic MAST-PP2A axis in breast cancer cells.

CIB2 and CIB3 are auxiliary subunits of the mechanotransduction channel of hair cells.

CIB2 is a Ca2+- and Mg2+-binding protein essential for mechanoelectrical transduction (MET) by cochlear hair cells, but not by vestibular hair cells that co-express CIB2 and CIB3. Here, we show that in cochlear hair cells, CIB3 can functionally substitute for CIB2. Using X-ray crystallography, we demonstrate that CIB2 and CIB3 are structurally similar to KChIP proteins, auxiliary subunits of voltage-gated Kv4 channels. CIB2 and CIB3 bind to TMC1/2 through a domain in TMC1/2 flanked by transmembrane domains 2 and 3. The co-crystal structure of the CIB-binding domain in TMC1 with CIB3 reveals that interactions are mediated through a conserved CIB hydrophobic groove, similar to KChIP1 binding of Kv4. Functional studies in mice show that CIB2 regulates TMC1/2 localization and function in hair cells, processes that are affected by deafness-causing CIB2 mutations. We conclude that CIB2 and CIB3 are MET channel auxiliary subunits with striking similarity to Kv4 channel auxiliary subunits.

TMIE Defines Pore and Gating Properties of the Mechanotransduction Channel of Mammalian Cochlear Hair Cells.

TMC1 and TMC2 (TMC1/2) have been proposed to form the pore of the mechanotransduction channel of cochlear hair cells. Here, we show that TMC1/2 cannot form mechanotransduction channels in cochlear hair cells without TMIE. TMIE binds to TMC1/2, and a TMIE mutation that perturbs TMC1/2 binding abolishes mechanotransduction. N-terminal TMIE deletions affect the response of the mechanotransduction channel to mechanical force. Similar to mechanically gated TREK channels, the C-terminal cytoplasmic TMIE domain contains charged amino acids that mediate binding to phospholipids, including PIP2. TMIE point mutations in the C terminus that are linked to deafness disrupt phospholipid binding, sensitize the channel to PIP2 depletion from hair cells, and alter the channel's unitary conductance and ion selectivity. We conclude that TMIE is a subunit of the cochlear mechanotransduction channel and that channel function is regulated by a phospholipid-sensing domain in TMIE with similarity to those in other mechanically gated ion channels.

Determination of the fate and biological responses of food additive silica particles in commercial foods.

Synthetic amorphous silica (SAS) is widely added to commercial foods as an anticaking agent. Concern about the potential application of nanosized silica in foods has increased as nanomaterials are not intended for use as food additives. This study evaluated the particle size distributions and biological responses of food additive SAS. An accurate, sensitive, and cost-effective analytical method for probing SAS was established, and quantitative analysis of its presence in commercial foods was performed. The results demonstrate that food additive SAS is an aggregated material composed of nanosized particles with nanosized aggregates of silica particles identified in commercial foods. Food additive SAS did not exhibit acute cytotoxicity compared to both general-grade nano (G-nano) and bulk (G-bulk) silica. Moreover, intestinal transport amounts of food additive SAS were significantly lower than for G-nano. Taken together, we find that food additive SAS does not exhibit acute toxicity resulting from nanosized materials.

Central Compensation in Auditory Brainstem after Damaging Noise Exposure.

Noise exposure is one of the most common causes of hearing loss and peripheral damage to the auditory system. A growing literature suggests that the auditory system can compensate for peripheral loss through increased central neural activity. The current study sought to investigate the link between noise exposure, increases in central gain, synaptic reorganization, and auditory function. All axons of the auditory nerve project to the cochlear nucleus, making it a requisite nucleus for sound detection. As the first synapse in the central auditory system, the cochlear nucleus is well positioned to respond plastically to loss of peripheral input. To investigate noise-induced compensation in the central auditory system, we measured auditory brainstem responses (ABRs) and auditory perception and collected tissue from mice exposed to broadband noise. Noise-exposed mice showed elevated ABR thresholds, reduced ABR wave 1 amplitudes, and spiral ganglion neuron loss. Despite peripheral damage, noise-exposed mice were hyperreactive to loud sounds and showed nearly normal behavioral sound detection thresholds. Ratios of late ABR peaks (2-4) relative to the first ABR peak indicated that brainstem pathways were hyperactive in noise-exposed mice, while anatomical analysis indicated there was an imbalance between expression of excitatory and inhibitory proteins in the ventral cochlear nucleus. The results of the current study suggest that a reorganization of excitation and inhibition in the ventral cochlear nucleus may drive hyperactivity in the central auditory system. This increase in central gain can compensate for peripheral loss to restore some aspects of auditory function.

Functional contributions of HCN channels in the primary auditory neurons of the mouse inner ear.

The hyperpolarization-activated current, Ih, is carried by members of the Hcn channel family and contributes to resting potential and firing properties in excitable cells of various systems, including the auditory system. Ih has been identified in spiral ganglion neurons (SGNs); however, its molecular correlates and their functional contributions have not been well characterized. To investigate the molecular composition of the channels that carry Ih in SGNs, we examined Hcn mRNA harvested from spiral ganglia of neonatal and adult mice using quantitative RT-PCR. The data indicate expression of Hcn1, Hcn2, and Hcn4 subunits in SGNs, with Hcn1 being the most highly expressed at both stages. To investigate the functional contributions of HCN subunits, we used the whole-cell, tight-seal technique to record from wild-type SGNs and those deficient in Hcn1, Hcn2, or both. We found that HCN1 is the most prominent subunit contributing to Ih in SGNs. Deletion of Hcn1 resulted in reduced conductance (Gh), slower activation kinetics (τfast), and hyperpolarized half-activation (V1/2) potentials. We demonstrate that Ih contributes to SGN function with depolarized resting potentials, depolarized sag and rebound potentials, accelerated rebound spikes after hyperpolarization, and minimized jitter in spike latency for small depolarizing stimuli. Auditory brainstem responses of Hcn1-deficient mice showed longer latencies, suggesting that HCN1-mediated Ih is critical for synchronized spike timing in SGNs. Together, our data indicate that Ih contributes to SGN membrane properties and plays a role in temporal aspects of signal transmission between the cochlea and the brain, which are critical for normal auditory function.

A Gata3-Mafb transcriptional network directs post-synaptic differentiation in synapses specialized for hearing.

Information flow through neural circuits is determined by the nature of the synapses linking the subtypes of neurons. How neurons acquire features distinct to each synapse remains unknown. We show that the transcription factor Mafb drives the formation of auditory ribbon synapses, which are specialized for rapid transmission from hair cells to spiral ganglion neurons (SGNs). Mafb acts in SGNs to drive differentiation of the large postsynaptic density (PSD) characteristic of the ribbon synapse. In Mafb mutant mice, SGNs fail to develop normal PSDs, leading to reduced synapse number and impaired auditory responses. Conversely, increased Mafb accelerates synaptogenesis. Moreover, Mafb is responsible for executing one branch of the SGN differentiation program orchestrated by the Gata3 transcriptional network. Remarkably, restoration of Mafb rescues the synapse defect in Gata3 mutants. Hence, Mafb is a powerful regulator of cell-type specific features of auditory synaptogenesis that offers a new entry point for treating hearing loss. DOI: http://dx.doi.org/10.7554/eLife.01341.001.

Explicit disassociation of a conditioned stimulus and unconditioned stimulus during extinction training reduces both time to asymptotic extinction and spontaneous recovery of a conditioned taste aversion.

Conditioned taste aversions (CTAs) may be acquired when an animal consumes a novel taste (CS) and then experiences the symptoms of poisoning (US). This aversion may be extinguished by repeated exposure to the CS alone. However, following a latency period in which the CS is not presented, the CTA will spontaneously recover (SR). In the current study we employed an explicitly unpaired extinction procedure (EU-EXT) to determine if it could thwart SR of a CTA. Sprague-Dawley rats acquired a strong CTA after 3 pairings of saccharin (SAC the CS) and Lithium Chloride (LiCl the US). CTA acquisition was followed by extinction (EXT) training consisting of either (a) CS-only exposure (CSO) or, (b) exposure to saccharin and Lithium Chloride on alternate days (i.e., explicitly unpaired: EU). Both extinction procedures resulted in >/= 90% reacceptance of SAC, although the EU extinction procedure (EU-EXT) significantly decreased the time necessary for rats to reach this criterion (compared to CSO controls). Rats were subsequently tested for SR of the CTA upon re-exposure to SAC following a 30-day latency period of water drinking. Rats that acquired a CTA and then underwent the CSO extinction procedure exhibited a significant suppression of SAC drinking during the SR test (as compared to their SAC drinking at the end of extinction). However, animals in the EU-EXT group did not show such suppression in drinking compared to CSO controls. These data suggest that the EU-EXT procedure may be useful in reducing both time to extinction and the spontaneous recovery of fears.