Engineering the Cellular Microenvironment: Integrating Three-Dimensional Nontopographical and Two-Dimensional Biochemical Cues for Precise Control of Cellular Behavior.

The development of biomaterials capable of regulating cellular processes and guiding cell fate decisions has broad implications in tissue engineering, regenerative medicine, and cell-based assays for drug development and disease modeling. Recent studies have shown that three-dimensional (3D) nanoscale physical cues such as nanotopography can modulate various cellular processes like adhesion and endocytosis by inducing nanoscale curvature on the plasma and nuclear membranes. Two-dimensional (2D) biochemical cues such as protein micropatterns can also regulate cell function and fate by controlling cellular geometries. Development of biomaterials with precise control over nanoscale physical and biochemical cues can significantly influence programming cell function and fate. In this study, we utilized a laser-assisted micropatterning technique to manipulate the 2D architectures of cells on 3D nanopillar platforms. We performed a comprehensive analysis of cellular and nuclear morphology and deformation on both nanopillar and flat substrates. Our findings demonstrate the precise engineering of single cell architectures through 2D micropatterning on nanopillar platforms. We show that the coupling between the nuclear and cell shape is disrupted on nanopillar surfaces compared to flat surfaces. Furthermore, our results suggest that cell elongation on nanopillars enhances nanopillar-induced endocytosis. We believe our platform serves as a versatile tool for further explorations into programming cell function and fate through combined physical cues that create nanoscale curvature on cell membranes and biochemical cues that control the geometry of the cell.

Pseudoneurotic Symptoms in the Schizophrenia Spectrum: A Longitudinal Study of Their Relation to Psychopathology and Clinical Outcomes.

BACKGROUND AND HYPOTHESIS: Nonpsychotic symptoms (depression, anxiety, obsessions, etc.) are frequent in schizophrenia-spectrum disorders and are usually conceptualized as comorbidity or transdiagnostic symptoms. However, in twentieth century foundational psychopathological literature, many nonpsychotic symptoms with specific phenomenology (here termed pseudoneurotic symptoms) were considered relatively typical of schizophrenia. In this prospective study, we investigated potential associations of pseudoneurotic symptoms with diagnostic status, functional outcome as well as psychopathological dimensions of schizophrenia. STUDY DESIGN: First-admitted patients (N = 121) diagnosed with non-affective psychosis, schizotypal disorder, or other mental illness were examined at initial hospitalization and 5 years later with a comprehensive assessment of psychopathology. Informed by the literature, we constructed scales targeting pseudoneurotic symptoms and other, more general, nonpsychotic symptoms. STUDY RESULTS: Pseudoneurotic symptoms aggregated in schizophrenia-spectrum groups compared to other mental illnesses and occurred at similar levels at baseline and follow-up. They longitudinally predicted poorer social and occupational functioning in schizophrenia-spectrum patients over a 5-year-period but not transition to schizophrenia-spectrum disorders from other mental illnesses. Finally, the level of pseudoneurotic symptoms correlated with disorder of basic self at both assessments and with positive and negative symptoms at follow-up. The scale targeting general nonpsychotic symptoms did not show this pattern of associations. CONCLUSIONS: The study supports that a group of nonpsychotic symptoms, ie, pseudoneurotic symptoms, are associated with schizophrenia-spectrum disorders and linked with temporally stable psychopathology, particularly disorder of the basic self. Their prospective association with social and occupational functioning needs replication.

Short-term biological variation of serum tryptase.

OBJECTIVES: Serum tryptase is a biomarker of mast cell activation. Among others, it is used in the diagnosis of anaphylaxis where a significant increase during the acute phase supports the diagnosis. When evaluating changes in biomarker levels, it is of utmost importance to consider the biological variation of the marker. Therefore, the aim of this study was to evaluate the short-term biological variation of serum tryptase. METHODS: Blood samples were drawn at 9 AM three days in a row from apparently healthy subjects. On day two, additional blood samples were drawn every third hour for 12 h. The tryptase concentration was measured in serum using a fluoroenzyme immunoassay (ImmunoCAP™, Thermo Fisher Scientific). Linear mixed-effects models were used to calculate components of biological variation. RESULTS: In 32 subjects, the overall mean concentration of tryptase was 4.0 ng/mL (range, 1.3-8.0 ng/mL). The within-subject variation was 3.7 % (95 % confidence interval (CI) 3.0-4.4 %), the between-subject variation was 31.5 % (95 % CI 23.1-39.8 %), and the analytical variation was 3.4 % (95 % CI 2.9-4.1 %). The reference change value was 13.3 % for an increase in tryptase at a 95 % level of significance. No significant day-to-day variation was observed (p=0.77), while a minute decrease in the serum concentration was observed during the day (p<0.0001). CONCLUSIONS: Serum tryptase is a tightly regulated biomarker with very low within-subject variation, no significant day-to-day variation, and only minor semidiurnal variation. In contrast, a considerable between-subject variation exists. This establishes serum tryptase as a well-suited biomarker for monitoring.

Extreme Reticulocytosis After Splenectomy in a Patient With Hemoglobin Mizuho.

BACKGROUND: Patients with Hb Mizuho may be splenectomized at a young age to decrease their need for blood transfusions. OBSERVATIONS: Transfusion-dependency decreased dramatically in a 4-year-old white boy with Hb Mizuho after splenectomy. Surprisingly, he developed reticulocytosis (>1000×10 9 /L) with a peak reticulocyte percentage of 49%, and erythrocyte abnormalities, including Heinz bodies, Howell-Jolly bodies, and basophilic stippling. Manual reticulocyte counting and flow cytometric measurement with anti-CD71 antibodies supported a truly elevated reticulocyte count. CONCLUSIONS: We propose possible explanations for the extreme reticulocytosis that arose postsplenectomy and compare the reticulocyte count in the present case with previously published cases.

Neointimal dissection - a rare complication to endovascular treatment in grafts and stent grafts.

BACKGROUND: Neointima formation and hyperplasia in vascular grafts may lead to graft complications threatening the patency of the vascular reconstruction. A rare complication to endovascular treatment of grafts and stent grafts is dissection inside the graft. CASE REPORT: We present here a case of a 69-year-old female with acute occlusion of the limb of an aorto-bifemoral graft for the third time, 16 years after the primary operation. As at the first two occasions, catheter-based intra-arterial thrombolysis was performed, but with residual stenosis inside the graft. During stent placement, dissection of the neointima or fibrin sheet occluded the inflow to the stent. The complication was resolved with placement of kissing stents. CONCLUSIONS: It is important to recognize iatrogenic neointima dissection inside graft and stent grafts, as continued thrombolysis will not solve this, but increase the risk of hemorrhagic complications.

Influence of age and sex on osmoscan indies for next-generation osmotic gradient ektacytometry.

BACKGROUND AND AIMS: Osmotic gradient ektacytometry is an important method for diagnosis of red blood cell membrane disorders. For interpretation of the osmoscan parameters on the ektacytomety, an age-matched control sample drawn at the same time is recommended for direct comparison. However, this can be challenging for laboratories to fulfil, especially when ektacytometry is performed in children. Therefore, the aim of this study was to evaluate the influence of age and sex on the osmoscan parameters. MATERIALS AND METHODS: Blood samples from 231 subjects were analyses on a LoRRca MaxSIS. Data were investigated for need of partitioning by age and sex. After outlier detection, reference intervals (RIs) for osmoscan parameters were estimated. RESULTS: For all parameters except EImin, lower values were observed in infants < 3 month (N = 50) than in all other age group. Hence, RIs were calculated separately for this age group. For EImin, a unified RI was calculated. No difference between sexes was observed for any of the parameters. CONCLUSION: Lower RIs and a left shift in the osmoscan curves were observed in infants < 3 months compared with older subjects. Hence, age-matched controls are necessary when evaluating ektacytometry in newborns, but can be ignored in older children and adults. This will ease the laboratory workflow when performing ektacytometry.

Norwegian radiologists' expectations of artificial intelligence in mammographic screening - A cross-sectional survey.

PURPOSE: To explore Norwegian breast radiologists' expectations of adding artificial intelligence (AI) in the interpretation procedure of screening mammograms. METHODS: All breast radiologists involved in interpretation of screening mammograms in BreastScreen Norway during 2021 and 2022 (n = 98) were invited to take part in this anonymous cross-sectional survey about use of AI in mammographic screening. The questionnaire included background information of the respondents, their expectations, considerations of biases, and ethical and social implications of implementing AI in screen reading. Data was collected digitally and analyzed using descriptive statistics. RESULTS: The response rate was 61% (60/98), and 67% (40/60) of the respondents were women. Sixty percent (36/60) reported ≥10 years' experience in screen reading, while 82% (49/60) reported no or limited experience with AI in health care. Eighty-two percent of the respondents were positive to explore AI in the interpretation procedure in mammographic screening. When used as decision support, 68% (41/60) expected AI to increase the radiologists' sensitivity for cancer detection. As potential challenges, 55% (33/60) reported lack of trust in the AI system and 45% (27/60) reported discrepancy between radiologists and AI systems as possible challenges. The risk of automation bias was considered high among 47% (28/60). Reduced time spent reading mammograms was rated as a potential benefit by 70% (42/60). CONCLUSION: The radiologists reported positive expectations of AI in the interpretation procedure of screening mammograms. Efforts to minimize the risk of automation bias and increase trust in the AI systems are important before and during future implementation of the tool.

Sub-threshold or full-syndrome borderline personality disorder in adolescents with recurrent self-harm - distinctly or dimensionally different?

BACKGROUND: Borderline personality disorder (BPD) is a severe mental disorder frequently seen in individuals with recurrent self-harm behaviour. To what extent there are distinguishing characteristics between self-harming adolescents who meet the criteria for a full diagnosis of BPD, a sub-threshold number of BPD criteria and those who don't have BPD, with respect to clinical characteristics, is still uncertain and could have important clinical implications. METHODS: Data from 103 adolescents with recurrent self-harm behaviour recruited from child and adolescent psychiatric outpatient clinics were collected through clinical interviews and self-reports. Bivariate analyses comparing participants with or without a diagnosis of BPD were performed. Group differences based on the number of BPD criteria fulfilled (few-if-any BPD: 0-2 criteria, sub-threshold BPD: 3-4 criteria, full-syndrome BPD: 5 or more criteria) were tested and regression analyses performed. RESULTS: Adolescents with a diagnosis of BPD (28.2%) had significantly higher numbers of co-morbid DSM-5 disorders, suicide attempts and self-harm methods. They also reported significantly higher levels of suicidal ideation, depression, anxiety and impulsivity, compared with adolescents without BPD. Adolescents with sub-threshold BPD (20.4%) place themselves in the intermediate position between participants with full-syndrome BPD and participants with few-if-any BPD, in terms of these symptoms. Higher levels of emotional regulation difficulties and a lower level of global functioning were significantly associated with fulfilling a higher number of BPD criteria. CONCLUSION: Adolescents with recurrent self-harm who meet diagnostic criteria for a full-syndrome BPD or sub-threshold BPD seem to have difficulties within the same spectrum. They seem dimensionally, but not categorically, different with respect to the severity of their difficulties. These adolescents need interventions aimed at their dysfunctional self-harm behaviour, emotional regulation difficulties and BPD symptoms at an earlier, rather than at a later stage of symptom development.

Pediatric reference intervals of the hemoglobin fractions HbA2, HbF and HbA0 using high-performance liquid chromatography and capillary electrophoresis.

INTRODUCTION: ß-thalassemia is a common genetic disorder with an estimated prevalence of 80-90 million carriers worldwide. As elevated hemoglobin A2 (HbA2) is a primary feature of carriers, hemoglobin fraction analysis is a common technique used for initial screening. However, pediatric reference intervals (RIs) are scarce. Hence, the aim was to establish pediatric RIs of hemoglobin fractions using high-performance liquid chromatography (HPLC) and capillary electrophoresis (CE). METHODS: Samples were collected from assumed healthy children and adolescents of 1-18 years. Analyses were conducted using the Tosoh Automated Glycohemoglobin Analyzer HLC-723®G11 (Tosoh G11, HPLC) and the Capillarys 3 Octa (CE). Data were investigated for need of partitioning by both age (1-6 years vs. 6-18 years) and sex. RESULTS: In total, 189 and 196 subjects were included in the statistical analysis of HPLC and CE, respectively. The 95% RI of HbA2 was 2.00-2.90% by HPLC and 2.2-3.0% by CE. Partitioning of data was not clinically relevant by HPLC. However, partitioning by age was suggested by CE. CONCLUSION: RIs of hemoglobin fractions in individuals of 1-18 years using commercially available HPLC and CE equipment were reported. This is the first report of a pediatric RI of HbA2 using the Tosoh G11.

Amyloid-like nanofibrous network confined and aligned ultrafine bimetallic nanozymes for smart antibacterial therapy.

Nanozyme-based antibacterial therapy (NABT) has emerged as a promising strategy to combat bacterial antimicrobial resistance. Engineering the noble metal nanozymes with strong bacterial capture and high catalytic activity for enhanced NABT is highly anticipated but still challenged. Herein, we developed hybrid nanozymes by engineering ultrafine bimetallic Au/Cu nanoparticles confined on the lysozyme amyloid-like nanofibrous networks (LNF). The introduction of copper in the nanozymes facilitates the H2O2 adsorption and reduces the energy barrier for activating the H2O2 decomposition to form •OH, meanwhile displaying the significantly enhanced POD-like activity under NIR irradiation. Taking advantage of the inherent supramolecular networks inspired from human defensin 6-trapping bacteria mechanism, the hybrid nanozymes effectively capture the bacteria and allow the catalytic attack around the bacterial surfaces to improve the antibacterial efficiency. Finally, the as-prepared nanozymes exhibit the preeminent bactericidal efficacy against bacteria, especially for drug-resistant bacteria both in vitro and in vivo, and the effect on wound healing.

In-situ PLL-g-PEG Functionalized Nanopore for Enhancing Protein Characterization.

Single-molecule nanopore detection technology has revolutionized proteomics research by enabling highly sensitive and label-free detection of individual proteins. Herein, we designed a small, portable, and leak-free flowcell made of PMMA for nanopore experiments. In addition, we developed an in situ functionalizing PLL-g-PEG approach to produce non-sticky nanopores for measuring the volume of diseases-relevant biomarker, such as the Alpha-1 antitrypsin (AAT) protein. The in situ functionalization method allows continuous monitoring, ensuring adequate functionalization, which can be directly used for translocation experiments. The functionalized nanopores exhibit improved characteristics, including an increased nanopore lifetime and enhanced translocation events of the AAT proteins. Furthermore, we demonstrated the reduction in the translocation event's dwell time, along with an increase in current blockade amplitudes and translocation numbers under different voltage stimuli. The study also successfully measures the single AAT protein volume (253 nm3 ), which closely aligns with the previously reported hydrodynamic volume. The real-time in situ PLL-g-PEG functionalizing method and the developed nanopore flowcell hold great promise for various nanopores applications involving non-sticky single-molecule characterization.

DSM-5 Non-Suicidal Self-Injury Disorder in a Clinical Sample of Adolescents with Recurrent Self-Harm Behavior.

INTRODUCTION: Non-suicidal self-injury disorder (NSSID) is a new diagnosis proposed in DSM-5 with a need of further study, especially in adolescent clinical populations where non-suicidal self-injury (NSSI) is particularly prevalent. We aimed to study characteristics of NSSID and estimate an optimal cutoff frequency level of NSSI behavior. METHODS: Data were collected from 103 outpatient adolescents (ages 12-18) with recurrent self-harm behavior. RESULTS: Adolescents with NSSID reported significantly more frequent NSSI behavior and suicide attempts than adolescents without NSSID. Frequency of NSSI, global functioning, depressive symptoms, number of self-harm methods and anxiety symptoms best discriminated between adolescents with and without NSSID. An optimal cutoff level for a diagnosis of NSSID was found to be ≥15 days with NSSI during the last year, which led to a reduction in the rate of adolescents diagnosed with NSSID from 54% to 46%. CONCLUSION: This study shows that NSSID is a highly impairing disorder characterized by high risk of multiple NSSI and suicide attempts, decreased functioning and other associated psychiatric disorders. Clinical awareness of these risks are important to ensure early detection and treatment. Future prospective longitudinal studies are needed to further validate the characteristics of the NSSID diagnosis and its clinical utility.

Short-term biological variation of plasma uracil in a Caucasian healthy population.

OBJECTIVES: Plasma uracil is a new biomarker to assess the activity of dihydropyrimidine dehydrogenase before cancer treatment with fluoropyrimidine drugs. Knowledge on the biological variation of plasma uracil is important to assess the applicability of plasma uracil as a biomarker of drug tolerance and efficacy. METHODS: A total of 33 apparently healthy individuals were submitted to sequential blood draws for three days. On the second day, blood draws were performed every third hour for 12 h. Plasma uracil was quantified by LC-MS/MS. The within-subject (CVI) and between-subject (CVG) biological variation estimates were calculated using linear mixed-effects models. RESULTS: The overall median value of plasma uracil was 10.6 ng/mL (range 5.6-23.1 ng/mL). The CVI and CVG were 13.5 and 22.1%, respectively. Plasma uracil remained stable during the day, and there was no day-to-day variation observed. No differences in biological variation components were found between sex and no correlation to age was found. Four samples were calculated to be required to estimate the homeostatic set-point ±15% with 95% confidence. CONCLUSIONS: Plasma uracil is subject to tight homeostatic regulation without semidiurnal and day-to-day variation, however between-subject variation exists. This emphasizes plasma uracil as a well-suited biomarker for evaluation of dihydropyrimidine dehydrogenase activity, but four samples are required to establish the homeostatic set-point in a patient.

Nursing students' clinical judgment skills in simulation and clinical placement: a comparison of student self-assessment and evaluator assessment.

BACKGROUND: Clinical judgment is an important and desirable learning outcome in nursing education. Students must be able to self-assess their clinical judgment in both the simulation and clinical settings to identify knowledge gaps and further improve and develop their skills. Further investigation is needed to determine the optimal conditions for and reliability of this self-assessment. AIMS: This study aimed to compare the same group of students' self-assessment of clinical judgment with an evaluator's assessment in both simulation and clinical settings. The study further aimed to investigate whether the Dunning-Kruger effect is present in nursing students' self-assessment of clinical judgment. METHODS: The study applied a quantitative comparative design. It was conducted in two learning settings: an academic simulation-based education course, and a clinical placement course in an acute care hospital. The sample consisted of 23 nursing students. The Lasater Clinical Judgment Rubric was used to collect data. The scores were compared using a t-test, intraclass correlation coefficient, Pearson's correlation coefficient, and Bland-Altman plots. The Dunning-Kruger effect was investigated using linear regression analysis and a scatter plot. RESULTS: The results showed an inconsistency between student self-assessment and evaluator assessment of clinical judgment in both simulation-based education and clinical placement. Students overestimated their clinical judgment when compared to the more experienced evaluator's assessment. Differences between students' scores and the evaluator's scores were larger when the evaluator's scores were low, indicating the presence of the Dunning-Kruger effect. CONCLUSION: It is vital to acknowledge that student self-assessment alone may not be a reliable predictor of a student's clinical judgment. Students who had a lower level of clinical judgment were likely to be less aware that this was the case. For future practice and research, we recommend a combination of student self-assessment and evaluator assessment to provide a more realistic view of students' clinical judgment skills.

A Validated Register-Based Algorithm to Identify Patients Diagnosed with Recurrence of Surgically Treated Stage I Lung Cancer in Denmark.

Introduction: Recurrence of cancer is not routinely registered in Danish national health registers. This study aimed to develop and validate a register-based algorithm to identify patients diagnosed with recurrent lung cancer and to estimate the accuracy of the identified diagnosis date. Material and Methods: Patients with early-stage lung cancer treated with surgery were included in the study. Recurrence indicators were diagnosis and procedure codes recorded in the Danish National Patient Register and pathology results recorded in the Danish National Pathology Register. Information from CT scans and medical records served as the gold standard to assess the accuracy of the algorithm. Results: The final population consisted of 217 patients; 72 (33%) had recurrence according to the gold standard. The median follow-up time since primary lung cancer diagnosis was 29 months (interquartile interval: 18-46). The algorithm for identifying a recurrence reached a sensitivity of 83.3% (95% CI: 72.7-91.1), a specificity of 93.8% (95% CI: 88.5-97.1), and a positive predictive value of 87.0% (95% CI: 76.7-93.9). The algorithm identified 70% of the recurrences within 60 days of the recurrence date registered by the gold standard method. The positive predictive value of the algorithm decreased to 70% when the algorithm was simulated in a population with a recurrence rate of 15%. Conclusion: The proposed algorithm demonstrated good performance in a population with 33% recurrences over a median of 29 months. It can be used to identify patients diagnosed with recurrent lung cancer, and it may be a valuable tool for future research in this field. However, a lower positive predictive value is seen when applying the algorithm in populations with low recurrence rates.

Impact of age-dependent red blood cell parameters on α-globin gene genotyping in children.

When screening for α-thalassemia in children, adult cut-offs for mean corpuscular volume (MCV) and mean corpuscular hemoglobin (MCH) are generally applied to guide genetic evaluation. However, the normal ranges for MCV and MCH are lower in children than in adults, so we hypothesized that using age-matched cut-offs could lead to a more rational diagnostic strategy. The aim of this study was to evaluate if age-matched cut-offs could be applied advantageously. Data on children referred to a hemoglobin fractionation at the Department of Clinical Biochemistry, Aarhus University Hospital between 2016-2021 were identified in the laboratory information system. α-globin gene (HBA1/HBA2) genotyping was performed using multiplex gap-polymerase chain reaction. A total of 387 children were identified. HBA1/HBA2-genotyping was performed in 207 children (53%), and α-thalassemia was diagnosed in 47 children (23%) with -α3.7/αα being the predominant genotype (13%). We found that 23 children had MCV and MCH levels in the normal age-matched range, and two of these children (9%) were α+ thalassemia carriers with three functional α-globin genes. Using age-specific cut-off levels resulted in a reduction of 23 (11%) genotypes performed. In conclusion, applying age-matched cut-offs for MCV and MCH when screening children for α-thalassemia lead to 11% fewer genotypes performed while 9% carriers of α+ thalassemia (of the medically innocuous genotype -α3.7/αα) would have been overlooked.

Pretreatment Platelet Count is a Prognostic Marker in Lung Cancer: A Danish Registry-based Cohort Study.

BACKGROUND: Thrombocytosis has been associated with a poor prognosis in a wide range of malignancies. However, the results have been conflicting for lung cancer. Therefore, we evaluated the prognostic value of platelet count in a large cohort of lung cancer patients. PATIENTS AND METHODS: All lung cancer patients diagnosed in The Central Denmark Region from 2009 to 2018 were included in the study. Data from the Danish Lung Cancer Registry were combined with data from the clinical laboratory information system on pretreatment platelet count. Platelet count was defined as low, normal, or high based on being below, within, or above the reference intervals. The prognostic value of platelet count was assessed by the Cox proportional hazard model. C-statistics were conducted to investigate if the platelet count added additional prognostic value to existing prognostic markers. RESULTS: Totally, 6,758 patients with non-small-cell lung cancer (NSCLC) and 1150 patients with small-cell lung cancer (SCLC) were included. Low and high platelet count were significantly associated with decreased overall survival (OS) in NSCLC patients (low: adjusted hazard ratio (HR)=1.75 (95% confidence interval [CI]: 1.49-2.06); high: adjusted HR=1.24 (95% CI: 1.16-1.33)). In SCLC patients, only low platelet count was significantly associated with decreased OS (adjusted HR = 2.71 [95% CI: 2.02-3.65]). C-statistics showed that the prognostic models were significantly improved by the addition of platelet count for both NSCLC and SCLC patients (P < .0001). CONCLUSION: Low and high platelet count were adverse prognostic factors in NSCLC patients, while only low platelet count was a prognostic marker in SCLC patients.

Exploring the NCS-382 Scaffold for CaMKIIα Modulation: Synthesis, Biochemical Pharmacology, and Biophysical Characterization of Ph-HTBA as a Novel High-Affinity Brain-Penetrant Stabilizer of the CaMKIIα Hub Domain.

Ca2+/calmodulin-dependent protein kinase II alpha (CaMKIIα) is a brain-relevant kinase and an emerging drug target for ischemic stroke and neurodegenerative disorders. Despite reported CaMKIIα inhibitors, their usefulness is limited by low subtype selectivity and brain permeability. (E)-2-(5-Hydroxy-5,7,8,9-tetrahydro-6H-benzo[7]annulen-6-ylidene)acetic acid (NCS-382) is structurally related to the proposed neuromodulator, γ-hydroxybutyric acid, and is a brain-penetrating high nanomolar-affinity ligand selective for the CaMKIIα hub domain. Herein, we report the first series of NCS-382 analogs displaying improved affinity and preserved brain permeability. Specifically, we present Ph-HTBA (1i) with enhanced mid-nanomolar affinity for the CaMKIIα binding site and a marked hub thermal stabilization effect along with a distinct CaMKIIα Trp403 flip upon binding. Moreover, Ph-HTBA has good cellular permeability and low microsomal clearance and shows brain permeability after systemic administration to mice, signified by a high Kp, uu value (0.85). Altogether, our study highlights Ph-HTBA as a promising candidate for CaMKIIα-associated pharmacological interventions and future clinical development.