Discovery of Pyrazolopyrazines as Selective, Potent, and Mutant-Active MET Inhibitors with Intracranial Efficacy.

Mesenchymal-epithelial transition factor (MET) is a receptor tyrosine kinase that serves a critical function in numerous developmental, morphogenic, and proliferative signaling pathways. If dysregulated, MET has been shown to be involved in the development and survival of several cancers, including non-small cell lung cancer (NSCLC), renal cancer, and other epithelial tumors. Currently, the clinical efficacy of FDA approved MET inhibitors is limited by on-target acquired resistance, dose-limiting toxicities, and less than optimal efficacy against brain metastasis. Therefore, there is still an unmet medical need for the development of MET inhibitors to address these issues. Herein we report the application of structure-based design for the discovery and development of a novel class of brain-penetrant MET inhibitors with enhanced activity against clinically relevant mutations and improved selectivity. Compound 13 with a MET D1228N cell line IC50 value of 23 nM showed good efficacy in an intracranial tumor model and increased the median overall survival of the animals to 100% when dosed orally at 100 mg/kg daily for 21 days.

Concurrent Validity of the Anxiety Disorders Section of the Anxiety Disorder Interview Schedule- Autism Spectrum Addendum (ADIS-ASA) in Autistic Youth.

PURPOSE: Examine the concurrent validity of specific Anxiety Disorders Section of the Anxiety Disorder Interview Schedule for DSM-IV-Autism Spectrum Addendum (ADIS-ASA)-Parent Interview in a sample of 167 autistic youth who met diagnostic criteria for an anxiety-related disorder (Mage = 9.91; 78.4% male; 82% non-Hispanic; 77.67% White). METHODS: Concurrent validity of Diagnostic and Statistical Manual (DSM)-defined ADIS-ASA anxiety disorder diagnostic caseness was examined via relations with (a) parent-reported dimensions of youth anxiety symptomology and (b) dimensional measures of youth anxiety-related functional impairment, respectively, using logistic regression models and point-biserial correlations. RESULTS: Significant relations were found between separation anxiety disorder and social anxiety disorder (but not generalized anxiety disorder nor obsessive-compulsive disorder) caseness, respectively, and theoretically consistent facets of dimensional youth anxiety symptomology. Relations between ADIS-ASA diagnostic caseness and youth functional impairment-related variables revealed that only separation anxiety disorder demonstrated robust evidence of convergent validity. CONCLUSION: Despite mixed findings concerning relations between ADIS-ASA anxiety disorder diagnostic caseness and dimensional measures of anxiety severity and anxiety-related impairment, the present findings provide further support for the status of the ADIS-ASA as a gold standard for assessment of anxiety in autistic youth. This work also highlights the importance of continuing to improve precision in measurement of anxiety symptomology in autistic youth, with implications for clinical assessment.

Fundamental processes in sensorimotor learning: Reasoning, refinement, and retrieval.

Motor learning is often viewed as a unitary process that operates outside of conscious awareness. This perspective has led to the development of sophisticated models designed to elucidate the mechanisms of implicit sensorimotor learning. In this review, we argue for a broader perspective, emphasizing the contribution of explicit strategies to sensorimotor learning tasks. Furthermore, we propose a theoretical framework for motor learning that consists of three fundamental processes: reasoning, the process of understanding action-outcome relationships; refinement, the process of optimizing sensorimotor and cognitive parameters to achieve motor goals; and retrieval, the process of inferring the context and recalling a control policy. We anticipate that this '3R' framework for understanding how complex movements are learned will open exciting avenues for future research at the intersection between cognition and action.

Differential effects of ganglioside lipids on the conformation and aggregation of islet amyloid polypeptide.

Despite causing over 1 million deaths annually, Type 2 Diabetes (T2D) currently has no curative treatments. Aggregation of the islet amyloid polypeptide (hIAPP) into amyloid plaques plays an important role in the pathophysiology of T2D and thus presents a target for therapeutic intervention. The mechanism by which hIAPP aggregates contribute to the development of T2D is unclear, but it is proposed to involve disruption of cellular membranes. However, nearly all research on hIAPP-lipid interactions has focused on anionic phospholipids, which are primarily present in the cytosolic face of plasma membranes. We seek here to characterize the effects of three gangliosides, the dominant anionic lipids in the outer leaflet of the plasma membrane, on the aggregation, structure, and toxicity of hIAPP. Our results show a dual behavior that depends on the molar ratio between the gangliosides and hIAPP. For each ganglioside, a low-lipid:peptide ratio enhances hIAPP aggregation and alters the morphology of hIAPP fibrils, while a high ratio eliminates aggregation and stabilizes an α-helix-rich hIAPP conformation. A more negative lipid charge more efficiently promotes aggregation, and a larger lipid headgroup improves inhibition of aggregation. hIAPP also alters the phase transitions of the lipids, favoring spherical micelles over larger tubular micelles. We discuss our results in the context of the available lipid surface area for hIAPP binding and speculate on a role for gangliosides in facilitating toxic hIAPP aggregation.

Does Local Aqueous Tobramycin Injection Reduce Open Fracture-Related Infection Rates?

OBJECTIVES: To examine the effect of local aqueous tobramycin injection adjunct to perioperative intravenous (IV) antibiotic prophylaxis in reducing fracture-related infections (FRIs) following reduction and internal fixation of open fractures. METHODS: Design: Retrospective cohort study. SETTING: Single academic Level I trauma center. PATIENT SELECTION CRITERIA: Patients with open extremity fractures treated via reduction and internal fixation with (intervention group) or without (control group) 80 mg of local aqueous (2mg/mL) tobramycin injected during closure at the time of definitive fixation were identified from December 2018 to August 2021 based upon population-matched demographic and injury characteristics. OUTCOME MEASURES AND COMPARISONS: The primary outcome was FRI within 6 months of definitive fixation. Secondary outcomes consisted of fracture nonunion and bacterial speciation. Differences in outcomes between the two groups were assessed and logistic regression models were created to assess the difference in infection rates between groups, with and without controlling for potential confounding variables, such as sex, fracture location, and Gustilo-Anderson classification. RESULTS: An analysis of 157 patients was performed with 78 patients in the intervention group and 79 patients in the control group. In the intervention group, 30 (38.5%) patients were female with mean age of 47.1 years. In the control group, 42 (53.2%) patients were female with mean age of 46.4 years. The FRI rate was 11.5% in the intervention group compared to 25.3% in the control group (p=0.026). After controlling for sex, Gustilo-Anderson classification, and fracture location, the difference in FRI rates between groups remained significantly different (p=0.014). CONCLUSIONS: Local aqueous tobramycin injection at the time of definitive internal fixation of open extremity fractures was associated with a significant reduction in fracture-related infection rates when administered as an adjunct to intravenous antibiotics, even after controlling for potential confounding variables. LEVEL OF EVIDENCE: Therapeutic Level III. See Instructions for Authors for a complete description of levels of evidence.

Unveiling Competitive Adsorption in TiO2 Photocatalysis through Machine-Learning-Accelerated Molecular Dynamics, DFT, and Experimental Methods.

The efficient harnessing of solar power for water treatment via photocatalytic processes has long been constrained by the challenge of understanding and optimizing the interactions at the photocatalyst surface, particularly in the presence of nontarget cosolutes. The adsorption of these cosolutes, such as natural organic matter, onto photocatalysts can inhibit the degradation of pollutants, drastically decreasing the photocatalytic efficiency. In the present work, computational methods are employed to predict the inhibitory action of a suite of small organic molecules during TiO2 photocatalytic degradation of para-chlorobenzoic acid (pCBA). Specifically, tryptophan, coniferyl alcohol, succinic acid, gallic acid, and trimesic acid were selected as interfering agents against pCBA to observe the resulting competitive reaction kinetics via bulk and surface phase reactions according to Langmuir-Hinshelwood adsorption dynamics. Experiments revealed that trimesic and gallic acids were most competitive with pCBA, followed by succinic acid. Density functional theory (DFT) and machine learning interatomic potentials (MLIPs) were used to investigate the molecular basis of these interactions. The computational findings showed that while the type of functional group did not directly predict adsorption affinity, the spatial arrangement and electronic interactions of these groups significantly influenced adsorption dynamics and corresponding inhibitory behavior. Notably, MLIPs, derived by fine-tuning models pretrained on a vastly larger dataset, enabled the exploration of adsorption behaviors over substantially longer periods than typically possible with conventional ab initio molecular dynamics, enhancing the depth of understanding of the dynamic interaction processes. Our study thus provides a pivotal foundation for advancing photocatalytic technology in environmental applications by demonstrating the critical role of molecular-level interactions in shaping photocatalytic outcomes.

Strong angular and spectral narrowing of electroluminescence in an integrated Tamm-plasmon-driven halide perovskite LED.

Next-generation light-emitting applications such as displays and optical communications require judicious control over emitted light, including intensity and angular dispersion. To date, this remains a challenge as conventional methods require cumbersome optics. Here, we report highly directional and enhanced electroluminescence from a solution-processed quasi-2-dimensional halide perovskite light-emitting diode by building a device architecture to exploit hybrid plasmonic-photonic Tamm plasmon modes. By exploiting the processing and bandgap tunability of the halide perovskite device layers, we construct the device stack to optimise both optical and charge-injection properties, leading to narrow forward electroluminescence with an angular full-width half-maximum of 36.6° compared with the conventional isotropic control device of 143.9°, and narrow electroluminescence spectral full-width half-maximum of 12.1 nm. The device design is versatile and tunable to work with emission lines covering the visible spectrum with desired directionality, thus providing a promising route to modular, inexpensive, and directional operating light-emitting devices.

Uncoupling histone modification crosstalk by engineering lysine demethylase LSD1.

Biochemical crosstalk between two or more histone modifications is often observed in epigenetic enzyme regulation, but its functional significance in cells has been difficult to discern. Previous enzymatic studies revealed that Lys14 acetylation of histone H3 can inhibit Lys4 demethylation by lysine-specific demethylase 1 (LSD1). In the present study, we engineered a mutant form of LSD1, Y391K, which renders the nucleosome demethylase activity of LSD1 insensitive to Lys14 acetylation. K562 cells with the Y391K LSD1 CRISPR knockin show decreased expression of a set of genes associated with cellular adhesion and myeloid leukocyte activation. Chromatin profiling revealed that the cis-regulatory regions of these silenced genes display a higher level of H3 Lys14 acetylation, and edited K562 cells show diminished H3 mono-methyl Lys4 near these silenced genes, consistent with a role for enhanced LSD1 demethylase activity. These findings illuminate the functional consequences of disconnecting histone modification crosstalk for a key epigenetic enzyme.

Optical and electronic functionality arising from controlled defect formation in nanoscale complex oxide lateral epitaxy.

Epitaxial crystallization of complex oxides provides the means to create materials with precisely selected composition, strain, and orientation, thereby controlling their functionalities. Extending this control to nanoscale three-dimensional geometries can be accomplished via a three-dimensional analog of oxide solid-phase epitaxy, lateral epitaxial crystallization. The orientation of crystals within laterally crystallized SrTiO3 systematically changes from the orientation of the SrTiO3 substrate. This evolution occurs as a function of lateral crystallization distance, with a rate of approximately 50° µm-1. The mechanism of the rotation is consistent with a steady-state stress of tens of megapascal over a 100-nanometer scale region near the moving amorphous/crystalline interface arising from the amorphous-crystalline density difference. Second harmonic generation and piezoelectric force microscopy reveal that the laterally crystallized SrTiO3 is noncentrosymmetric and develops a switchable piezoelectric response at room temperature, illustrating the potential to use lateral crystallization to control the functionality of complex oxides.

Anterior Vertebral Body Tethering: A Single-Center Cohort with 4.3 to 7.4 Years of Follow-up.

BACKGROUND: Vertebral body tethering (VBT) is a well-recognized, non-fusion alternative for idiopathic scoliosis in children with growth remaining. To date, there have been almost no published outcome studies with postoperative follow-up of >2 years. We aimed to fill this gap by evaluating mid-term outcomes in our first 31 consecutive patients. METHODS: We retrospectively assessed additional clinical and radiographic data (mean, 5.7 ± 0.7 years) from our first 31 consecutive patients. Assessments included standard deformity measures, skeletal maturity status, and any additional complications (e.g., suspected broken tethers or surgical revisions). Using the same definition of success (i.e., all residual deformities, instrumented or uninstrumented, ≤30° at maturity; no posterior spinal fusion), we revisited the success rate, revision rate, and suspected broken tether rate. RESULTS: Of our first 31 patients treated with VBT, 29 (of whom 28 were non-Hispanic White and 1 was non-Hispanic Asian; 27 were female and 2 were male) returned for additional follow-up. The success rate dropped to 64% with longer follow-up as deformity measures increased, and the revision rate increased to 24% following 2 additional surgical revisions. Four additional suspected broken tethers were identified, for a rate of 55%, with only 1 occurring beyond 4 years. No additional patients had conversion to a posterior spinal fusion. We observed a mean increase of 4° (range, 2° to 8°) in main thoracic deformity measures and 8° (range, 6° to 12°) in thoracolumbar deformity measures. CONCLUSIONS: With >5 years of follow-up, we observed a decrease in postoperative success, as progression of the deformity was observed in most subgroups, and an increase in the revision and suspected broken tether rates. No additional patients had conversion to a posterior spinal fusion, which may indicate long-term survivorship. LEVEL OF EVIDENCE: Therapeutic Level III. See Instructions for Authors for a complete description of levels of evidence.

A prospective pilot study of gut microbiome in cerebral vasospasm and delayed cerebral ischemia after aneurysmal subarachnoid hemorrhage.

A recent systematic review indicated that gut-microbiota-brain axis contributes to growth and rupture of intracranial aneurysms. However, gaps were detected in the role of intestinal microbiome in cerebral vasospasm (CVS) after aneurysmal subarachnoid hemorrhage (aSAH). This is the first pilot study aiming to test study feasibility and identify differences in gut microbiota between subjects with and without CVS following aSAH. A prospective nested case-control pilot study with 1:1 matching was conducted recruiting subjects with aSAH: cases with CVS; and controls without CVS based on the clinical picture and structured bedside transcranial Doppler (TCD). Fecal samples for microbiota analyses by means of 16S rRNA gene amplicon sequencing were collected within the first 96 h after ictus. Operational taxonomic unit tables were constructed, diversity metrics calculated, phylogenetic trees built, and differential abundance analysis (DAA) performed. At baseline, the groups did not differ significantly in basic demographic and aneurysm-related characteristics (p > 0.05). Alpha-diversity (richness and Shannon Index) was significantly reduced in cases of middle cerebral artery (MCA) vasospasm (p < 0.05). In DAA, relative abundance of genus Acidaminococcus was associated with MCA vasospasm (p = 0.00013). Two butyrate-producing genera, Intestinimonas and Butyricimonas, as well as [Clostridium] innocuum group had the strongest negative correlation with the mean blood flow velocity in anterior cerebral arteries (p < 0.01; rho = - 0.63; - 0.57, and - 0.57, respectively). In total, 16 gut microbial genera were identified to correlate with TCD parameters, and two intestinal genera correlated with outcome upon discharge. In this pilot study, we prove study feasibility and present the first preliminary evidence of gut microbiome signature associating with CVS as a significant cause of stroke in subjects with aSAH.

Maintenance of germline stem cell homeostasis despite severe nuclear distortion.

Stem cell loss in aging and disease is associated with nuclear deformation. Yet, how nuclear shape influences stem cell homeostasis is poorly understood. We investigated this connection using Drosophila germline stem cells, as survival of these stem cells is compromised by dysfunction of the nuclear lamina, the extensive protein network that lines the inner nuclear membrane and gives shape to the nucleus. To induce nuclear distortion in germline stem cells, we used the GAL4-UAS system to increase expression of the permanently farnesylated nuclear lamina protein, Kugelkern, a rate limiting factor for nuclear growth. We show that elevated Kugelkern levels cause severe nuclear distortion in germline stem cells, including extensive thickening and lobulation of the nuclear envelope and nuclear lamina, as well as alteration of internal nuclear compartments. Despite these changes, germline stem cell number, proliferation, and female fertility are preserved, even as females age. Collectively, these data demonstrate that disruption of nuclear architecture does not cause a failure of germline stem cell survival or homeostasis, revealing that nuclear deformation does not invariably promote stem cell loss.

Deep learning of Parkinson's movement from video, without human-defined measures.

BACKGROUND: The core clinical sign of Parkinson's disease (PD) is bradykinesia, for which a standard test is finger tapping: the clinician observes a person repetitively tap finger and thumb together. That requires an expert eye, a scarce resource, and even experts show variability and inaccuracy. Existing applications of technology to finger tapping reduce the tapping signal to one-dimensional measures, with researcher-defined features derived from those measures. OBJECTIVES: (1) To apply a deep learning neural network directly to video of finger tapping, without human-defined measures/features, and determine classification accuracy for idiopathic PD versus controls. (2) To visualise the features learned by the model. METHODS: 152 smartphone videos of 10s finger tapping were collected from 40 people with PD and 37 controls. We down-sampled pixel dimensions and videos were split into 1 s clips. A 3D convolutional neural network was trained on these clips. RESULTS: For discriminating PD from controls, our model showed training accuracy 0.91, and test accuracy 0.69, with test precision 0.73, test recall 0.76 and test AUROC 0.76. We also report class activation maps for the five most predictive features. These show the spatial and temporal sections of video upon which the network focuses attention to make a prediction, including an apparent dropping thumb movement distinct for the PD group. CONCLUSIONS: A deep learning neural network can be applied directly to standard video of finger tapping, to distinguish PD from controls, without a requirement to extract a one-dimensional signal from the video, or pre-define tapping features.

Scalable Generation of 3D Pancreatic Islet Organoids from Human Pluripotent Stem Cells in Suspension Bioreactors.

We describe a scalable method for the robust generation of 3D pancreatic islet-like organoids from human pluripotent stem cells using suspension bioreactors. Our protocol involves a 6-stage, 20-day directed differentiation process, resulting in the production of 104-105 organoids. These organoids comprise α- and ß-like cells that exhibit glucose-responsive insulin and glucagon secretion. We detail methods for culturing, passaging, and cryopreserving stem cells as suspended clusters and for differentiating them through specific growth media and exogenous factors added in a stepwise manner. Additionally, we address quality control measures, troubleshooting strategies, and functional assays for research applications.

Non-Alcoholic Fatty Liver Disease Induced by Feeding Medium-Chain Fatty Acids Upregulates Cholesterol and Lipid Homeostatic Genes in Skeletal Muscle of Neonatal Pigs.

Non-alcoholic fatty liver disease (NAFLD) is a range of disorders characterized by lipid accumulation in hepatocytes. Although this spectrum of disorders is associated with adult obesity, recent evidence suggests that this condition could also occur independently of obesity, even in children. Previously, we reported that pigs fed a formula containing medium-chain fatty acids (MCFAs) developed hepatic steatosis and weighed less than those fed an isocaloric formula containing long-chain fatty acids (LCFAs). Our objective was to determine the association between NAFLD and the skeletal muscle transcriptome in response to energy and lipid intake. Neonatal pigs were fed one of three formulas: a control formula (CONT, n = 6) or one of two isocaloric high-energy formulas containing either long (LCFA, n = 6) or medium (MCFA, n = 6) chain fatty acids. Pigs were fed for 22 d, and tissues were collected. Body weight at 20 and 22 d was greater for LCFA-fed pigs than their CONT or MCFA counterparts (p < 0.005). Longissimus dorsi weight was greater for LCFA compared with MCFA, while CONT was intermediate (p < 0.05). Lean gain and protein deposition were greater for LCFA than for CONT and MCFA groups (p < 0.01). Transcriptomic analysis revealed 36 differentially expressed genes (DEGs) between MCFA and LCFA, 53 DEGs between MCFA and CONT, and 52 DEGs between LCFA and CONT (FDR < 0.2). Feeding formula high in MCFAs resulted in lower body and muscle weights. Transcriptomics data suggest that the reduction in growth was associated with a disruption in cholesterol metabolism in skeletal muscles.

Randomized Controlled Trial of Irrigation-Coupled Bipolar Electrocautery Versus Tourniquet in Total Knee Arthroplasty.

Background: Recovery from total knee arthroplasty remains arduous for some patients, prompting interest in perioperative management. While tourniquet use is not associated with longer-term outcomes, its effect on quadriceps strength in the immediate postoperative window is unknown. Methods: A single-center, double-blind, randomized controlled trial of 66 patients undergoing primary total knee arthroplasty from 2019 to 2022 was performed to compare the use of an irrigation-coupled bipolar device (ICBD) and no tourniquet (ICBD group, N = 34) to tourniquet use with no ICBD (tourniquet group, N = 32). Groups were similar with respect to age, sex, and obesity. The primary outcome was quadriceps strength at 2 weeks, measured using a handheld dynamometer and standardized to the contralateral side. Knee Injury and Osteoarthritis Outcome Score for Joint Replacement was measured with the difference from baseline serving as a secondary outcome. Comparisons were performed using the Student's t-test. Results: Only 28 patients, 14 in each group, had primary outcome data. At 2-weeks, quadriceps strength was higher in the ICBD group compared to the tourniquet group (83% vs 70%), though not statistically significant (P = .16). There was no difference between the ICBD and tourniquet groups in Knee Injury and Osteoarthritis Outcome Score for Joint Replacement changed at 2-weeks (13 vs 10, P = .37) or 6-weeks (16 vs 17, P = .76). Conclusions: Tourniquet use was associated with a small but not statistically significant difference in quadriceps strength at 2 weeks that may justify further study given the loss of power here. There can be limitations to conducting randomized controlled trials that are important for early-career investigators to consider and that were magnified due to COVID-related restrictions in the present study, which we discuss. Level of Evidence: Level II.

Endovascular Treatment of Basilar Apex Aneurysms: An Updated Systematic Review and Meta-Analysis in the Era of Flow Diversion.

BACKGROUND: Endovascular options for the treatment of basilar apex aneurysms (BAAs) are heterogeneous, and evidence is limited to retrospective cohorts and case series. We seek to evaluate the efficacy and complications associated with various endovascular treatment methods of BAAs. METHODS: Systematic review of PubMed, Embase, and Web of Science adhering to the PRISMA guidelines. Retrospective and prospective studies evaluating endovascular treatment of BAAs between January 2010 and July 2024 were included. Relevant information including occlusion rates, aneurysm recurrence, retreatment rates, and complications were subjected to meta-analysis. RESULTS: Fifteen studies with 1,049 BAAs were included. The median aneurysm diameter was 8.5 mm (range 4.6-19.75), with a median follow-up of 33.7 months (range 6.0-117.6). Residual aneurysm filling occurred in 24% after primary coiling (95% CI=0.16-0.32), 25% after single stent-assisted coiling (s-SAC; 95% CI=0.04-0.46), 25% after Y-stenting (95% CI=0.12-0.37), and 23% after flow diverter stenting (FDS; 95% CI=0.11-0.35). Recurrence rates were high for primary coiling (27%, 95% CI=0.18-0.36) and s-SAC (19%, 95% CI=0.13-0.26), but significantly lower for Y-stenting (9%, 95% CI=0.03-0.15) and FDS (4%, 95% CI=-0.04-0.11). Retreatment rates were 19% for primary coiling (95% CI=0.12-0.26), 17% for s-SAC (95% CI=0.07-0.27), 5% for Y-stenting (95% CI=-0.03-0.12), and 13% for FDS (95% CI=-0.01-0.27). Meta-regression indicated larger aneurysms had higher complication rates (p=0.02). Thromboembolic events were most frequent with FDS and Y-stenting (12%). CONCLUSION: Occlusion rates were similar across treatments, but recurrence rates were significantly lower after Y-stenting and FDS treatments compared to primary coiling, although they carried a higher number of thromboembolic complications.