Candesartan restores blood-brain barrier dysfunction, mitigates aberrant gene expression, and extends lifespan in a knockin mouse model of epileptogenesis.

Blockade of Angiotensin type 1 receptor (AT1R) has potential therapeutic utility in the treatment of numerous detrimental consequences of epileptogenesis, including oxidative stress, neuroinflammation, blood-brain barrier (BBB) dysfunction. We have recently shown that many of these pathological processes played a critical role in seizure onset and propagation in the Scn8a-N1768D mouse model. Here we investigate the efficacy and potential mechanism(s) of action of candesartan (CND), an FDA-approved angiotensin receptor blocker (ARB) indicated for hypertension, in improving outcomes in this model of pediatric epilepsy. We compared length of lifespan, seizure frequency, and BBB permeability in juvenile (D/D) and adult (D/+) mice treated with CND at times after seizure onset. We performed RNAseq on hippocampal tissue to quantify differences in genome-wide patterns of transcript abundance and inferred beneficial and detrimental effects of canonical pathways identified by enrichment methods in untreated and treated mice. Our results demonstrate that treatment with CND gives rise to increased survival, longer periods of seizure freedom, and diminished BBB permeability, as well as partially reversed or "normalized" disease-induced genome-wide gene expression profiles associated with inhibition of NF-κB, TNFα, IL-6, and TGF-ß signaling in juvenile and adult mice. Pathway analyses reveal that efficacy of CND is due to its known dual mechanism of action as both an AT1R antagonist and a PPARγ agonist. The robust efficacy of CND across ages, sexes and mouse strains is a positive indication for its translation to humans and its suitability of use for clinical trials in children with SCN8A epilepsy.

Disease-modifying rdHSV-CA8* non-opioid analgesic gene therapy treats chronic osteoarthritis pain by activating Kv7 voltage-gated potassium channels.

Chronic pain is common in our population, and most of these patients are inadequately treated, making the development of safer analgesics a high priority. Knee osteoarthritis (OA) is a primary cause of chronic pain and disability worldwide, and lower extremity OA is a major contributor to loss of quality-adjusted life-years. In this study we tested the hypothesis that a novel JDNI8 replication-defective herpes simplex-1 viral vector (rdHSV) incorporating a modified carbonic anhydrase-8 transgene (CA8*) produces analgesia and treats monoiodoacetate-induced (MIA) chronic knee pain due to OA. We observed transduction of lumbar DRG sensory neurons with these viral constructs (vHCA8*) (~40% of advillin-positive cells and ~ 50% of TrkA-positive cells colocalized with V5-positive cells) using the intra-articular (IA) knee joint (KJ) route of administration. vHCA8* inhibited chronic mechanical OA knee pain induced by MIA was dose- and time-dependent. Mechanical thresholds returned to Baseline by D17 after IA KJ vHCA8* treatment, and exceeded Baseline (analgesia) through D65, whereas negative controls failed to reach Baseline responses. Weight-bearing and automated voluntary wheel running were improved by vHCA8*, but not negative controls. Kv7 voltage-gated potassium channel-specific inhibitor XE-991 reversed vHCA8*-induced analgesia. Using IHC, IA KJ of vHCA8* activated DRG Kv7 channels via dephosphorylation, but negative controls failed to impact Kv7 channels. XE-991 stimulated Kv7.2-7.5 and Kv7.3 phosphorylation using western blotting of differentiated SH-SY5Y cells, which was inhibited by vHCA8* but not by negative controls. The observed prolonged dose-dependent therapeutic effects of IA KJ administration of vHCA8* on MIA-induced chronic KJ pain due to OA is consistent with the specific activation of Kv7 channels in small DRG sensory neurons. Together, these data demonstrate for the first-time local IA KJ administration of vHCA8* produces opioid-independent analgesia in this MIA-induced OA chronic pain model, supporting further therapeutic development.

Generation of Marfan syndrome-specific induced pluripotent stem cells harboring FBN1 mutations.

Marfan syndrome (MFS) is a hereditary condition caused by mutations in the FBN1 gene. Genetic mutations in the FBN1 locus impact the function of the encoded protein, Fibrillin 1, a structural molecule forming microfibrils found in the connective tissue. MFS patients develop severe cardiovascular complications including thoracic aortic aneurysm and aortic dissection, which predispose them to an enhanced risk of premature death. Here, we generated two induced pluripotent stem cell (iPSC) lines harboring mutations in the FBN1 gene (p.C1942C>A and c.1954 T>C), directly derived from MFS patients. We have shown that both iPSC lines displayed expression of pluripotency markers, normal karyotype and ability of trilineage differentiation, representing a valuable tool for the identification of new therapeutic strategies for intervening in this disease.

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NADPH oxidase 2 activity disrupts Calmodulin/CaMKIIα complex via redox modifications of CaMKIIα-contained Cys30 and Cys289: Implications in Parkinson's disease.

Ca2+/calmodulin-dependent protein kinase II α (CaMKIIα) signaling in the brain plays a critical role in regulating neuronal Ca2+ homeostasis. Its dysfunctional activity is associated with various neurological and neurodegenerative disorders, including Parkinson's disease (PD). Using computational modeling analysis, we predicted that, two essential cysteine residues contained in CaMKIIα, Cys30 and Cys289, may undergo redox modifications impacting the proper functioning of the CaMKIIα docking site for Ca2+/CaM, thus impeding the formation of the CaMKIIα:Ca2+/CaM complex, essential for a proper modulation of CaMKIIα kinase activity. Our subsequent in vitro investigations confirmed the computational predictions, specifically implicating Cys30 and Cys289 residues in impairing CaMKIIα:Ca2+/CaM interaction. We observed CaMKIIα:Ca2+/CaM complex disruption in dopamine (DA) nigrostriatal neurons of post-mortem Parkinson's disease (PD) patients' specimens, addressing the high relevance of this event in the disease. CaMKIIα:Ca2+/CaM complex disruption was also observed in both in vitro and in vivo rotenone models of PD, where this phenomenon was associated with CaMKIIα kinase hyperactivity. Moreover, we observed that, NADPH oxidase 2 (NOX2), a major enzymatic generator of superoxide anion (O2●-) and hydrogen peroxide (H2O2) in the brain with implications in PD pathogenesis, is responsible for CaMKIIα:Ca2+/CaM complex disruption associated to a stable Ca2+CAM-independent CaMKIIα kinase activity and intracellular Ca2+ accumulation. The present study highlights the importance of oxidative stress, in disturbing the delicate balance of CaMKIIα signaling in calcium dysregulation, offering novel insights into PD pathogenesis.

Assessing ChatGPT's Competency in Addressing Interdisciplinary Inquiry on Chatbot Uses in Sports Rehabilitation: Descriptive Analysis.

BACKGROUND: ChatGPT showcases exceptional conversational capabilities and extensive cross-disciplinary knowledge. In addition, it can perform multiple roles in a single chat session. This unique multi-role-playing feature positions ChatGPT as a promising tool for exploring interdisciplinary subjects. OBJECTIVE: The study intended to evaluate ChatGPT's competency in addressing interdisciplinary inquiry, using a case study on exploring the opportunities and challenges of chatbot uses in sports rehabilitation. METHODS: We developed a model termed PanelGPT to assess ChatGPT's competency in addressing interdisciplinary topics through simulated panel discussions. Taking chatbot uses in sports rehabilitation -an interdisciplinary topic -as an example, we prompted ChatGPT through PanelGPT to role-play a physiotherapist, psychologist, nutritionist, AI expert, and an athlete in a simulated panel discussion. During the simulation, we posed questions to the panel, while ChatGPT acted as both the panelists for responses and the moderator for steering the discussion. We performed the simulation using ChatGPT-4 and evaluated the responses by referring to literature and our human expertise. RESULTS: By tackling questions related to chatbot uses in sport rehabilitation on patient education, physiotherapy, physiology, nutrition, and ethical consideration, responses from the ChatGPT-simulated panel discussion reasonably pointed to the benefits such as 24/7 support, personalized advice, automated tracking, and reminders. It also correctly emphasized the importance of patient education and identified challenges such as limited interaction modes, inaccuracies in emotion-related advice, assurance of data privacy and security, transparency in data handling, and fairness in model training. It also stressed that chatbots are to assist as a co-pilot, not to replace human healthcare professionals in the rehabilitation process. CONCLUSIONS: ChatGPT exhibits strong competency in addressing interdisciplinary inquiry by simulating multiple experts from complementary backgrounds, with significant implications in assisting medical education. CLINICALTRIAL: Not applicable.

Improved tumour delivery of iron oxide nanoparticles for magnetic hyperthermia therapy of melanoma via ultrasound guidance and 111In SPECT quantification.

Magnetic field hyperthermia relies on the intra-tumoural delivery of magnetic nanoparticles by interstitial injection, followed by their heating on exposure to a remotely-applied alternating magnetic field (AMF). This offers a potential sole or adjuvant route to treating drug-resistant tumours for which no alternatives are currently available. However, two challenges in nanoparticle delivery currently hinder the effective clinical translation of this technology: obtaining enough magnetic material within the tumour to enable sufficient heating; and doing this accurately to limit or avoid damage to surrounding healthy tissue. A further complication is the lack of established methods to non-invasively quantify nanoparticle biodistribution, which is necessary to evaluate the performance of improved delivery strategies. Here we employ 111In radiolabelling and single-photon emission computed tomography (SPECT) to non-invasively quantify distribution of a clinical grade iron-oxide-based nanoparticle in a mouse model of melanoma. We show that compared to manual injection, ultrasound guided delivery together with syringe-pump-controlled infusion improves both the nanoparticle concentration within the tumour, and the accuracy of delivery - reducing off-target peri-tumoural delivery. Following AMF heating, injected melanomas shrank significantly compared to non-injected controls, validating therapeutic efficacy. Systemic off-target delivery was quantified and extrapolated to predict off-target energy absorbance within safe limits for the main sites of background accumulation. With many nanoparticle-based therapies currently in development for cancer, this image-guided delivery strategy has wide potential impact beyond the field of magnetic hyperthermia. Future use in representative patient cohorts would also be enabled by the high clinical availability of both SPECT and ultrasound imaging.

Hypertension and Ventilatory Responses During Exercise in the Fitness Registry and the Importance of Exercise National Database (FRIEND).

BACKGROUND: A high minute ventilation/rate of carbon dioxide production (V̇E/V̇co2) slope during exercise is prognostic for cardiovascular death. Recent data indicate that adults with either controlled or untreated primary hypertension, but not those with uncontrolled hypertension, exhibit a higher V̇E/V̇co2 slope during exercise. However, the sample sizes were modest. Therefore, we used the Fitness Registry and the Importance of Exercise National Database to determine whether adults with hypertension, particularly those with controlled or untreated hypertension, exhibit higher V̇E/V̇co2 slopes compared with adults without hypertension. METHODS AND RESULTS: Using the Fitness Registry and the Importance of Exercise National Database, we isolated primary hypertension by excluding those with any disease other than hypertension or taking any medications other than antihypertension medications. We also excluded current smokers and those with obesity. The V̇E/V̇co2 slope was determined during a peak cycling exercise test. All data are presented as median [interquartile range]. We compared groups using linear regression adjusted for age, male/female, and body mass index. The characteristics of the entire sample (n=4109) were age, 42 [18] years; 48% women; body mass index, 26 [4] kg/m2. The V̇E/V̇co2 slope did not differ between adults with hypertension (n=1940; 24.7 [3.7]) compared with those without hypertension (n=2169; 24.9 [3.8]) (hypertension versus no hypertension, P=0.31; overall model: R2=0.07, F4,4104=73.0; P<0.001). Further, the V̇E/V̇co2 slope did not differ between adults with medication-controlled hypertension (n=107; 24.4 [3.0]), untreated hypertension (n=1626; 24.8 [3.9]), uncontrolled hypertension (n=207; 24.8 [3.0]), or those without hypertension (n=2169; 24.9 [3.8]) (hypertension subgroup versus no hypertension, P≥0.06; overall model: R2=0.07, F6,4102=49.6, P<0.001). CONCLUSIONS: Primary hypertension is not associated with V̇E/V̇co2 slope in the Fitness Registry and the Importance of Exercise National Database.

EMpowerment of PArents in THe Intensive Care: A multicentre validation study in Japan.

BACKGROUND: The importance of assessing family satisfaction in paediatric intensive care units (PICUs) is becoming increasingly recognised. The survey, EMpowerment of Parents in THe Intensive Care "EMPATHIC-30", was designed to assess family satisfaction and has been translated and implemented in several countries but not yet in Japan. OBJECTIVES: The objective of this study was to translate, culturally adapt, and validate the EMPATHIC-30 questionnaire in Japanese and to identify potential factors for family-centred care satisfaction. METHODS: We translated and adapted for patient-reported outcome measures via a 10-step process outlined by the Principles of Good Practice. Four paediatric PICUs in Japan participated in the validation study, and the parental enrolment criterion was a child with a PICU stay of >24 h. Reliability was measured by Cronbach's α, and congruent validity was tested with overall satisfaction-with-care scales by correlation analysis. Multivariate linear regression modelling was conducted to identify factors related to each domain of the Japanese EMPATHIC-30. RESULTS: A total of 163 parents (mean age: 31.9 ± 5.4 years; 81% were mothers) participated. The five domains of the Japanese EMPATHIC-30 showed high reliability (α = 0.87 to 0.97) and congruent validity, demonstrating high correlations with overall satisfaction in nurses (r = 0.75) and doctors (r = 0.76). Multivariate modelling found that elective admission, mechanical ventilation, and parents who had experience of a family member in an adult intensive care unit had higher satisfaction scores in all five domains (p < 0.05). Moreover, Buddhists assigned higher satisfaction scores in the Care and Treatment domain (p = 0.03). CONCLUSIONS: The Japanese EMPATHIC-30 questionnaire has demonstrated adequate reliability and validity measures. We also identified that elective admission, mechanical ventilation, and having previous adult intensive care unit experience of a family member were factors in assigning higher scores for all satisfaction domains. PICU clinicians need to be cognisant of ethical, cultural, and religious factors relating to the critically ill child and their family.

Formation of a Pseudoaneurysm after a .22 Caliber Bullet Embolized to the Left Femoral Artery: A Case Report.

Bullet embolization is a rare and potentially life-threatening complication of gunshot wounds, particularly in lowpowered and small-caliber bullets. When these small bullets enter a large elastic vessel, they have the potential to leave a small entrance hole that can form a traumatic pseudoaneurysm. These pseudoaneurysms, which may be life-protecting at first, may rupture and lead to exsanguination if not found. We report an interesting case of an 18-year-old male gunshot victim where a bullet formed an aortic pseudoaneurysm and subsequently embolized and present a review of the literature regarding bullet embolization and traumatic pseudoaneurysms.

Geographic and demographic trends with the initiation of virtual interviews in general surgery: A 7-program evaluation.

BACKGROUND: Although most general surgery residency interviews remain virtual, the effect of this format remains understudied. Single-institution data have shown an increase in the number of applications received and interviews conducted with virtual interviewing but no change in the geographic backgrounds of interviewed or matched applicants. This study sought to compare national trends in geographic characteristics of general surgery applicants, interviewed applicants, and matched applicants between in-person and virtual application cycles. STUDY DESIGN: A retrospective review of 7 general surgery residency programs from application years 2016-2019 (in-person) and 2020-2021 (virtual) was conducted. Data collected included birth year, sex, race, medical school state, and contact location at the time of application. Data were analyzed using generalized mixed effects linear models. RESULTS: A total of 52,742 applicants, 4,550 interviewed applicants, and 329 matched applicants were included. During virtual application cycles, there were no increases in the average number of applicants (P = .25), interviewed applicants (P = .36), or matched (P = .84) applicants per year. Virtual cycles were associated with a larger proportion of interviews conducted with applicants from out-of-state medical schools (P < .01) and listing out-of-state contact locations (P < .01) compared with in-person application cycles. There were no significant geographic differences in matched applicants between virtual and in-person application cycles. CONCLUSION: Virtual application cycles had greater geographic diversity among interviewed applicants. However, similar differences were not seen in the geographic diversity of matched applicants. Additional efforts should focus on why no changes in the geographic diversity of matched applicants were identified.

Rapid Microbial Profiling through Multimodal Biosensors for Transversal Analysis.

The intricate interactions between host and microbial communities hold significant implications for biology and medicine. However, traditional microbial profiling methods face limitations in processing time, measurement of absolute abundance, detection of low biomass, discrimination between live and dead cells, and functional analysis. This study introduces a rapid multimodal microbial characterization platform, Multimodal Biosensors for Transversal Analysis (MBioTA), for capturing the taxonomy, viability, and functional genes of the microbiota. The platform incorporates single cell biosensors, scalable microwell arrays, and automated image processing for rapid transversal analysis in as few as 2 h. The multimodal biosensors simultaneously characterize the taxon, viability, and functional gene expression of individual cells. By automating the image processing workflow, the single cell analysis techniques enable the quantification of bacteria with sensitivity down to 0.0075%, showcasing its capability in detecting low biomass samples. We illustrate the applicability of the MBioTA platform through the transversal analysis of the gut microbiota composition, viability, and functionality in a familial Alzheimer's disease mouse model. The effectiveness, rapid turnaround, and scalability of the MBioTA platform will facilitate its application from basic research to clinical diagnostics, potentially revolutionizing our understanding and management of diseases associated with microbe-host interactions.

Membrane Curvature Promotes ER-PM Contact Formation via Junctophilin-EHD Interactions.

Contact sites between the endoplasmic reticulum (ER) and the plasma membrane (PM) play a crucial role in governing calcium regulation and lipid homeostasis. Despite their significance, the factors regulating their spatial distribution on the PM remain elusive. Inspired by observations in cardiomyocytes, where ER-PM contact sites concentrate on tubular PM invaginations known as transverse tubules (T-tubules), we hypothesize that the PM curvature plays a role in ER-PM contact formation. Through precise control of PM invaginations, we show that PM curvatures locally induce the formation of ER-PM contacts in cardiomyocytes. Intriguingly, the junctophilin family of ER-PM tethering proteins, specifically expressed in excitable cells, is the key player in this process, while the ubiquitously expressed extended synaptotagmin 2 does not show a preference for PM curvature. At the mechanistic level, we find that the low complexity region (LCR) and the MORN motifs of junctophilins can independently bind to the PM, but both the LCR and MORN motifs are required for targeting PM curvatures. By examining the junctophilin interactome, we identify a family of curvature-sensing proteins, Eps15-homology domain containing proteins (EHDs), that interact with the MORN_LCR motifs and facilitate junctophilins' preferential tethering to curved PM. These findings highlight the pivotal role of PM curvature in the formation of ER-PM contacts in cardiomyocytes and unveil a novel mechanism for the spatial regulation of ER-PM contacts through PM curvature modulation.

Patient Interest in Quadriceps Autograft Anterior Cruciate Ligament Reconstruction Is Increasing Over Other Autograft Options: A 12-Year Google Trends Analysis.

Purpose: To use Google trends to explore differences in public interest among types of anterior cruciate ligament (ACL) autografts, specifically quadriceps tendon, patellar tendon, and hamstring tendon autografts, between 2008 and 2019. Methods: Data were obtained by querying Google Trends for key terms and phrases for online search data ranging from January 2008 to December 2019. Relative search volumes were created based on searches related to ACL reconstruction with comparative analysis generated for search terms related to quadriceps ACL, patellar tendon ACL, and hamstring ACL autografts. Statistical analysis included linear regression analysis, comparison of quarterly search volume trends over time, and comparison of cumulative annual search volumes for 2008 versus 2019. Results: Linear models for respective search terms were statistically significant for the quadriceps (P < .001) and patellar (P = .007) tendon autograft groups but not the hamstring group (P = .129). The quadriceps autograft group demonstrated a 12-year search volume trend change of 0.56, which was significantly greater than the hamstring (0.07; P < .001) and patellar tendon (0.168; P < .001) groups. There was no significant difference in the trend change between hamstring and patellar tendon groups (P = .20). Percent change in cumulative relative annual search volumes between 2008 and 2019 was 112% for the quadriceps tendon group, 12.9% for the hamstring group, and 18.6% for the patellar tendon group. Conclusions: This study indicates a consistently increasing public interest in quadriceps tendon autograft for ACL reconstruction. The quadriceps autograft group demonstrated a significantly greater 12-year online search volume, greater linear correlation, and larger percent change between 2008 and 2019 compared with patellar tendon or hamstring autograft groups. Clinical Relevance: Awareness of patient perceptions has value in informing shared decision-making, aligning patient expectations, and guiding areas of future research. Each of these has an impact on patient care. Being aware of patient interest and expectations is particularly important in areas with controversial or emerging research.

Arthroscopic Posterior Capsular Release Improves Range of Motion and Outcomes for Flexion Contracture After Anterior Cruciate Ligament Reconstruction in Athletes.

Purpose: To assess outcomes of arthroscopic posterior capsular release among athletes for loss of terminal extension following anterior cruciate ligament (ACL) reconstruction. Methods: A retrospective review of prospectively collected data was performed for patients undergoing arthroscopic posterior capsular release for knee extension loss following ACL reconstruction between January 2014 and December 2019. Procedure indications included extension loss greater than 10° at least 3 months after ACL reconstruction that was refractory to physical therapy. Patients were included if they were involved in either high school or college athletics, had complete outcomes of interest, and had at least 2 years of follow-up. Prospectively collected outcomes included preoperative and postoperative measurement of knee extension, International Knee Documentation Committee score, Lysholm score, return to sport data, and complications. Results: Eighteen athletes with minimum 2 years of follow-up who underwent posterior capsular release following ACL reconstruction performed by a single surgeon were included in the analysis. Patients underwent surgery at an average of 16 weeks after ACL reconstruction. Knee extension improved an average of 13.8° at 2 years' follow-up (prerelease mean extension deficit 15.1°, postrelease mean extension deficit 1.3°, P < .005). Improvements in the International Knee Documentation Committee score averaged 21.7 at 6 months and 35.0 at 24 months, both of which were statistically significant (P < .001). Similarly, differences in Lysholm included a significant improvement of 23.0 and 34.2 at 6 months and 2 years, respectively (P < .001). In total, 77.8% returned to sport at an average of 9.8 months from their primary ACL surgery and 6.5 months following posterior capsular release surgery. No infections or neurovascular complications were observed. One patient required secondary release to achieve adequate extension. Conclusions: For athletes with persistent knee extension loss after ACL reconstruction, knee extension was significantly improved at 2 years following arthroscopic posterior capsular release. Substantial improvements in patient-reported outcomes also were seen. In addition, subjects demonstrated a high rate of return to sport and return to preinjury performance levels. Level of Evidence: Level IV, therapeutic case series.