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Single-stranded DNA (ssDNA) templates along with Cas9 have been used for knocking-in exogenous sequences in the genome but suffer from low efficiency. Here, we show that ssDNA with chemical modifications in 12-19% of internal bases, which we denote as enhanced ssDNA (esDNA), improve knock-in (KI) by 2-3-fold compared to end-modified ssDNA in airway basal stem cells (ABCs), CD34 + hematopoietic cells (CD34 + cells), T-cells and endothelial cells. Over 50% of alleles showed KI in three clinically relevant loci (CFTR, HBB and CCR5) in ABCs using esDNA and up to 70% of alleles showed KI in the HBB locus in CD34 + cells in the presence of a DNA-PKcs inhibitor. This level of correction is therapeutically relevant and is comparable to adeno-associated virus-based templates. The esDNA templates did not improve KI in induced pluripotent stem cells (iPSCs). This may be due to the absence of the nuclease TREX1 in iPSCs. Indeed, knocking out TREX1 in other cells improved KI using unmodified ssDNA. esDNA can be used to modify 20-30 bp regions in primary cells for therapeutic applications and biological modeling. The use of this approach for gene length insertions will require new methods to produce long chemically modified ssDNA in scalable quantities., (© The Author(s) 2024. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Background: Clinical prediction models for pediatric community-acquired pneumonia (CAP) may standardize management. Understanding physician risk thresholds is important for model implementation. We aimed to elucidate physician-derived thresholds for chest radiograph performance and empirical antibiotic treatment of CAP among children presenting to the emergency department with respiratory illness before and after knowledge of results of a validated clinical prediction model., Methods: Pediatric emergency physicians were surveyed through the American Academy of Pediatrics Pediatric Emergency Medicine Collaborative Research Committee and provided 8 clinical vignettes for children with respiratory symptoms. Respondents were asked to indicate their probability of radiographic CAP and choose whether they would obtain a chest radiograph or give empirical antibiotics before and after being provided with the probability of radiographic CAP based on a validated prediction model. We used logistic regression to establish testing and treatment thresholds, defined as the disease probability at which half of physicians acted., Results: Two-hundred and eight (44.3%) of 469 physicians completed the survey. Most were attending physicians (96.0%) practicing in a freestanding children's hospital (76.8%). Testing and treatment thresholds for CAP were 17.6% (95% confidence interval [CI] 16.4% to 18.8%) and 66.1% (95% CI 60.1% to 72.5%), respectively, before knowledge of the model-estimated probability. With knowledge of the prediction model, testing and treatment thresholds were 13.5% (95% CI 12.3% to 14.7%) and 58.0% (95% CI 53.2-62.8)., Conclusions: We elucidated physician thresholds for testing and treatment of CAP, which may be integrated into future pneumonia risk models to improve acceptability and incorporation into practice., Competing Interests: CONFLICT OF INTEREST DISCLOSURES: The authors have indicated they have no conflicts of interest relevant to this article to disclose., (Copyright © 2024 by the American Academy of Pediatrics.)

Objective: Postoperative pain is the most common undesirable outcome after neurosurgery. Ketorolac is a nonsteroidal anti-inflammatory drug (NSAID) that is administered parenterally and carries a theoretical increased risk of bleeding. Our study aims to determine whether ketorolac after craniotomy for tumor resection significantly changes the rate of postoperative adverse events, adequately controls pain, and decreases concurrent narcotic use., Methods: We performed a retrospective chart review of all adult patients who underwent craniotomy for brain tumor resection from 2013 to 2022. Analysis of patients who received ketorolac and those who did not in the postoperative period were compared for adverse events associated with ketorolac use. Secondary outcomes included patient-reported pain scores and postoperative opioid use., Results: In total, 1114 patients were included, of whom 70 received ketorolac in the postoperative period. Ketorolac was typically administered to patients in whom narcotics had failed to provide sufficient pain relief. Patients receiving ketorolac were younger (P = 0.001) and had a lower comorbidity index (P = 0.041) compared with the nonketorolac group. Patients receiving ketorolac did not experience a significantly increased rate of bleeding events (P = 0.850). Patients receiving ketorolac had significantly greater baseline levels of pain (P = 0.018) and opioid use (P = 0.047). When matched for chronic comorbidities including pain disorders, the ketorolac group only displayed greater levels of pain early in the postoperative course (postoperative day 0-1) but not in latter part of the initial postoperative period., Conclusions: Ketorolac is a safe and effective option for pain control after craniotomy for tumor resection. Prospective data are needed to better validate these retrospective observations., (Copyright © 2024 Elsevier Inc. All rights reserved.)

The major genetic risk factor for Alzheimer's disease (AD), APOE4, accelerates beta-amyloid (Aβ) plaque formation, but whether this is caused by APOE expressed in microglia or astrocytes is debated. We express here the human APOE isoforms in astrocytes in an Apoe-deficient AD mouse model. This is not only sufficient to restore the amyloid plaque pathology but also induces the characteristic transcriptional pathological responses in Apoe-deficient microglia surrounding the plaques. We find that both APOE4 and the protective APOE2 from astrocytes increase fibrillar plaque deposition, but differentially affect soluble Aβ aggregates. Microglia and astrocytes show specific alterations in function of APOE genotype expressed in astrocytes. Our experiments indicate a central role of the astrocytes in APOE mediated amyloid plaque pathology and in the induction of associated microglia responses., Competing Interests: Disclosure and competing interests statement BDS is or has been a consultant for Eli Lilly, Biogen, Janssen Pharmaceutica, Eisai, AbbVie and other companies. BDS is an Editorial Board Member for EMBO Molecular Medicine. BDS is also a scientific founder of Augustine Therapeutics and a scientific founder and stockholder of Muna Therapeutics., (© 2024. The Author(s).)

Autonomous laboratories can accelerate discoveries in chemical synthesis, but this requires automated measurements coupled with reliable decision-making 1,2 . Most autonomous laboratories involve bespoke automated equipment 3-6 , and reaction outcomes are often assessed using a single, hard-wired characterization technique 7 . Any decision-making algorithms 8 must then operate using this narrow range of characterization data 9,10 . By contrast, manual experiments tend to draw on a wider range of instruments to characterize reaction products, and decisions are rarely taken based on one measurement alone. Here we show that a synthesis laboratory can be integrated into an autonomous laboratory by using mobile robots 11-13 that operate equipment and make decisions in a human-like way. Our modular workflow combines mobile robots, an automated synthesis platform, a liquid chromatography-mass spectrometer and a benchtop nuclear magnetic resonance spectrometer. This allows robots to share existing laboratory equipment with human researchers without monopolizing it or requiring extensive redesign. A heuristic decision-maker processes the orthogonal measurement data, selecting successful reactions to take forward and automatically checking the reproducibility of any screening hits. We exemplify this approach in the three areas of structural diversification chemistry, supramolecular host-guest chemistry and photochemical synthesis. This strategy is particularly suited to exploratory chemistry that can yield multiple potential products, as for supramolecular assemblies, where we also extend the method to an autonomous function assay by evaluating host-guest binding properties., (© 2024. The Author(s).)

Extended reality (XR) technologies are emerging as promising platforms for medical training and procedural guidance, particularly in complex cardiac interventions. This paper presents a high-fidelity methodology to perform real-time 3D catheter tracking and visualization during simulated cardiac interventions. A custom 3D-printed setup with mounted cameras enables biplane video capture of a catheter. A computer vision algorithm processes the biplane images in real-time to reconstruct the 3D catheter trajectory represented by any designated number of points along its length. This method accurately localizes the catheter tip within 1 mm and can reconstruct any arbitrary catheter configuration. The tracked catheter data is integrated into an interactive Unity-based scene rendered on the Meta Quest 3 headset. The visualization seamlessly combines a reconstructed 3D patient-specific heart model with the dynamically tracked catheter, creating an immersive extended reality training environment. Our experimental study, involving six participants, demonstrated that the 3D visualization provided by the proposed XR system significantly outperformed 2D visualization in terms of speed and user experience. This suggests that the XR system has the potential to enhance catheterization training by improving spatial comprehension and procedural skills. The proposed system demonstrates the potential of XR technologies to transform percutaneous cardiac interventions through improved visualization and interactivity., (© 2024. The Author(s).)

Objective: To examine perceived OR ergonomics facilitators and barriers, with a focus on the interdisciplinary team., Summary Background Data: Poor ergonomics causes musculoskeletal injuries affecting all operating room (OR) staff with repercussions on patient care, outcomes, and sustainability. Lack of ergonomic awareness and education are risk factors., Methods: We conducted a self-administered web-based survey of OR nurses, surgeons, and anesthesiologists at a single centre (n=238). We developed a questionnaire through items generation and reduction, followed by reliability and validity testing., Results: Response rate was 53.8%. Respondents perceived that on average 80% of nurses, 70% of surgeons, and 40% anesthesiologists experienced MSK injuries, with no difference in professional groups' perceptions. Guideline ergonomics interventions were rarely used (<25%) except for specialized clothing (33%), equipment repositioning (59%), and seating (37%), though perceived beneficial by 80-90%. Reported barriers to optimal ergonomics were organizational/structural (lack of time, space, equipment, funding), whereas solutions were individual. Fear of unfavourable perception from others was a concern for 62%. Teams discussing, prioritizing, monitoring, or helping with ergonomics was indicated by <50%. Individual ergonomic adaptations were perceived as convenience by other staff., Conclusions: While structural/organizational issues are reported as barriers to ergonomics, solutions appeared as individual responsibilities. Team dynamics did not prioritize nor support ergonomics. Education tools leveraging the interdisciplinary team are warranted. This work will be supplemented by interviews and live observations to build tailored educational tools for OR teams., Competing Interests: Conflicts of interest and source of funding: None to declare. Funding was received from the Sunnybrook AFP-SHARE Fund as peer-reviewed operating grant., (Copyright © 2024 Wolters Kluwer Health, Inc. All rights reserved.)

Neural stem cells have intact innate immune responses that protect them from virus infection and cell death. Yet, viruses can antagonize such responses to establish neuropathogenesis. Using a forebrain organoid model system at two developmental time points, we identified that neural stem cells, in particular radial glia, are basally primed to respond to virus infection by upregulating several antiviral interferon-stimulated genes. Infection of these organoids with a neuropathogenic Enterovirus-D68 strain, demonstrated the ability of this virus to impede immune activation by blocking interferon responses. Together, our data highlight immune gene signatures present in different types of neural stem cells and differential viral capacity to block neural-specific immune induction., (© 2024. The Author(s).)

Importance: Health datasets from clinical sources do not reflect the breadth and diversity of disease, impacting research, medical education, and artificial intelligence tool development. Assessments of novel crowdsourcing methods to create health datasets are needed., Objective: To evaluate if web search advertisements (ads) are effective at creating a diverse and representative dermatology image dataset., Design, Setting, and Participants: This prospective observational survey study, conducted from March to November 2023, used Google Search ads to invite internet users in the US to contribute images of dermatology conditions with demographic and symptom information to the Skin Condition Image Network (SCIN) open access dataset. Ads were displayed against dermatology-related search queries on mobile devices, inviting contributions from adults after a digital informed consent process. Contributions were filtered for image safety and measures were taken to protect privacy. Data analysis occurred January to February 2024., Exposure: Dermatologist condition labels as well as estimated Fitzpatrick Skin Type (eFST) and estimated Monk Skin Tone (eMST) labels., Main Outcomes and Measures: The primary metrics of interest were the number, quality, demographic diversity, and distribution of clinical conditions in the crowdsourced contributions. Spearman rank order correlation was used for all correlation analyses, and the χ2 test was used to analyze differences between SCIN contributor demographics and the US census., Results: In total, 5749 submissions were received, with a median of 22 (14-30) per day. Of these, 5631 (97.9%) were genuine images of dermatological conditions. Among contributors with self-reported demographic information, female contributors (1732 of 2596 contributors [66.7%]) and younger contributors (1329 of 2556 contributors [52.0%] aged <40 years) had a higher representation in the dataset compared with the US population. Of 2614 contributors who reported race and ethnicity, 852 (32.6%) reported a racial or ethnic identity other than White. Dermatologist confidence in assigning a differential diagnosis increased with the number of self-reported demographic and skin-condition-related variables (Spearman R = 0.1537; P < .001). Of 4019 contributions reporting duration since onset, 2170 (54.0%) reported onset within less than 7 days of submission. Of the 2835 contributions that could be assigned a dermatological differential diagnosis, 2523 (89.0%) were allergic, infectious, or inflammatory conditions. eFST and eMST distributions reflected the geographical origin of the dataset., Conclusions and Relevance: The findings of this survey study suggest that search ads are effective at crowdsourcing dermatology images and could therefore be a useful method to create health datasets. The SCIN dataset bridges important gaps in the availability of images of common, short-duration skin conditions.

Murine neonatal neutrophil depletion strategies have problems achieving deep neutrophil clearance and accurate residual neutrophil fraction detection. An isotype switch method can achieve profound neutrophil clearance using a combination of anti-Ly6G and anti-rat κ Ig light chain antibodies in adult C57Bl/6 mice, proven by extra- and intracellular Ly6G detection by flow cytometry. We adapted this technique to neonatal mice, testing four neutrophil depletion strategies in the peripheral circulation, bone marrow, and spleen. Four protocols were tested: P3 Ly6G and P1-3 Ly6G (anti-Ly6G on postnatal days (P) 3 and 1-3 respectively), and P3 Dual and P1-3 Dual (anti-Ly6G and anti-rat κ Ig light chain on P3 and P1-3 respectively). Intracellular and extracellular Ly6G presence was detected using flow cytometry. Isotype control antibodies were used as controls. P1-3 Dual protocol achieved significantly better neutrophil depletion than the P1-3 Ly6G or P3 Ly6G protocols (97% vs. 74% and 97% vs. 50%, respectively) in the peripheral circulation. The P3 Dual protocol alone was enough to achieve significantly better neutrophil clearance (93%) than any of the Ly6G alone protocols. The Ly6G alone protocols led to near-total elimination of extracellular Ly6G. However, there was a significant presence of intracellular Ly6G in the CD45+ cell population, evading detection by extracellular Ly6G antibody-based detection methods. P3 protocols perform better than P1-3 protocols for bone marrow and splenic neutrophil clearance. Thus, the P3 Dual protocol might be the most effective and ethical protocol to induce profound neutrophil depletion in neonatal mice, an alternative to daily anti-Ly6G injections., Summary Sentence: Dual antibody-based neutrophil depletion effectively induces circulating neutrophil clearance in neonatal mice.

A series of donor-acceptor (D-π-A) substituted diphenylbutadienes exhibiting solvatochromic emission and a large Stokes shift (100-200 nm) were designed and synthesized for distinctive organelle labelling, enabling real-time monitoring of organelle behaviour such as lysosomal dynamics, mitophagy monitoring, and stress responses. The morpholine-substituted D-A-D diphenylbutadiene (M2) was employed to investigate selective imaging of lysosomes, the uptake of damaged mitochondria through mitophagy, and monitoring lysosomal viscosity or pH changes. Other diphenylbutadiene derivatives (M1, M3, M4) selectively accumulated in lipid droplets. All the synthesized derivatives demonstrated significant uptake in 5-day post-fertilization zebrafish larvae, with M2 showing maximum uptake in the enterocyte-containing heart and intestinal regions, which include the lysosomes., (© 2024 Wiley-VCH GmbH.)

Human subcutaneous adipose tissue (SAT) contains a diverse array of cell-types; however, the epigenomic landscape among the SAT cell-types has remained elusive. Our integrative analysis of single-cell resolution DNA methylation and chromatin conformation profiles (snm3C-seq), coupled with matching RNA expression (snRNA-seq), systematically cataloged the epigenomic, 3D topology, and transcriptomic dynamics across the SAT cell-types. We discovered that the SAT CG methylation (mCG) landscape is characterized by pronounced hyper-methylation in myeloid cells and hypo-methylation in adipocytes and adipose stem and progenitor cells (ASPCs), driving nearly half of the 705,063 detected differentially methylated regions (DMRs). In addition to the enriched cell-type-specific transcription factor binding motifs, we identified TET1 and DNMT3A as plausible candidates for regulating cell-type level mCG profiles. Furthermore, we observed that global mCG profiles closely correspond to SAT lineage, which is also reflected in cell-type-specific chromosome compartmentalization. Adipocytes, in particular, display significantly more short-range chromosomal interactions, facilitating the formation of complex local 3D genomic structures that regulate downstream transcriptomic activity, including those associated with adipogenesis. Finally, we discovered that variants in cell-type level DMRs and A compartments significantly predict and are enriched for variance explained in abdominal obesity. Together, our multimodal study characterizes human SAT epigenomic landscape at the cell-type resolution and links partitioned polygenic risk of abdominal obesity to SAT epigenome.

Objectives: Hospitalizations for children with medical complexity (CMC) have increased substantially over the past 2 decades and constitute a disproportionate percentage of hospitalization rates and costs among children. We sought to describe the etiology and utilization for hospitalizations of CMC using the Pediatric Clinical Classification System (PECCS)., Methods: Using the 2019 Kids' Inpatient Database, we classified hospitalizations for CMC using the PECCS, which groups diagnoses into mutually exclusive, pediatric-specific categories. For the medical, surgical, and medical/surgical PECCS clinical groups, we reported diagnosis groups accounting for ≥1% of hospital encounters for that group. We described admission frequency, cost, payer, length of stay, and mortality rates within each diagnosis grouping using survey-weighted statistics., Results: We identified 2 315 743 nonlivebirth hospitalizations, of which 712 139 (30.8%) were for CMC. Most (94.4%) hospitalizations occurred at a teaching hospital. Medical diagnosis comprised most hospitalizations (69.2%), whereas hospitalizations for surgical and medical/surgical conditions had a higher median cost. The most common diagnosis groups overall were encounters for chemotherapy, diabetic ketoacidosis, and respiratory failure, whereas the costliest were for necrotizing enterocolitis, transposition of the great vessels, and hypoplastic left heart syndrome., Conclusions: We evaluated the most common diagnoses and their associated resource use for hospitalized CMC using the PECCS, providing a more granular view on the etiology, utilization, cost, and outcomes of hospitalizations for CMC. These topics represent high-impact areas for further research and quality efforts for CMC., (Copyright © 2024 by the American Academy of Pediatrics.)

Background: The MeMed BV Test produces a score that increases with increasing likelihood of bacterial infection. We evaluated its association with radiographic pneumonia in children., Methods: We performed a secondary analysis of a multicenter prospective study of febrile children 90 days to 18 years presenting to an emergency department. We evaluated the association of the MeMed BV test with radiographic pneumonia in adjusted logistic regression models., Results: Of 182 children, 74 (41%) had radiographic pneumonia. Among children with a high likelihood of having viral illness per the BV test, 26% had radiographic pneumonia; this increased to 64% among those with a BV test which indicated a high likelihood of bacterial infection. The sensitivity and specificity for radiographic pneumonia when using a BV test classification of moderate or high likelihood of bacterial infection were 60.8% and 62.0%, respectively. A BV test indicating the highest likelihood of bacterial infection had 23.61 higher adjusted odds (95% confidence interval 6.30-88.6) of radiographic pneumonia. The most common radiographic finding among children classified as having a high likelihood of viral infection by the BV test was interstitial opacities. The most common finding among children classified as having a high likelihood of bacterial infection were infiltrates. Except for antibiotic use, clinical outcomes occurred in similar proportions by BV category., Conclusions: Children with moderate to high likelihoods of bacterial infection on the BV test had higher odds of radiographic pneumonia. Apart from antibiotic use, the test was not significantly associated with clinical outcomes in this study., (Copyright © 2024 by the American Academy of Pediatrics.)

Understanding matrix molecular activities that regulate the postnatal growth and remodeling of the temporomandibular joint (TMJ) articular disc and condylar cartilage will enable the development of effective regenerative strategies targeting TMJ disorders. This study elucidated the distinct roles of type V collagen (collagen V) in regulating these two units. Studying the TMJ of young adult Col5a1 +/- mice, we found that loss of collagen V resulted in substantial changes in the proliferation, clustering and density of progenitors in condylar cartilage, but did not have a major impact on disc cells that are more fibroblast-like. Although loss of collagen V led to thickened collagen fibrils with increased heterogeneity in the disc, there were no significant changes in local micromodulus, except for a reduction at the posterior end of the inferior side. Following the induction of aberrant occlusal loading by the unilateral anterior crossbite (UAC) procedure, both wild-type (WT) and Col5a1 +/- condylar cartilage exhibited salient remodeling, and Col5a1 +/- condyle developed more pronounced degeneration and tissue hypertrophy at the posterior end than the WT. In contrast, neither UAC nor collagen V deficiency induced marked changes in the morphology or biomechanical properties of the disc. Together, our findings highlight the distinct roles of collagen V in regulating these two units during postnatal growth and remodeling, emphasizing its more crucial role in condylar cartilage due to its impact on the highly mechanosensitive progenitors. These results provide the foundation for using collagen V to improve the regeneration of TMJ and the care of patients with TMJ disorders. STATEMENT OF SIGNIFICANCE: Successful regeneration of the temporomandibular joint (TMJ) articular disc and condylar cartilage remains a significant challenge due to the limited understanding of matrix molecular activities that regulate the formation and remodeling of these tissues. This study demonstrates that collagen V plays distinct and critical roles in these processes. In condylar cartilage, collagen V is essential for regulating progenitor cell fate and maintaining matrix integrity. In the disc, collagen V also regulates fibril structure and local micromechanics, but has a limited impact on cell phenotype or its remodeling response. Our findings establish collagen V as a key component in maintaining the integrity of these two units, with a more crucial role in condylar cartilage due to its impact on progenitor cell activities., Competing Interests: Declaration of competing interests The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Effective river water quality monitoring is essential for sustainable water resource management. In this study, we established a comprehensive monitoring system along the Kaveri River, capturing real-time data on multiple critical water quality parameters. The parameters collected encompassed water contamination levels, turbidity, pH measurements, temperature, and total dissolved solids (TDS), providing a holistic view of river water quality. The monitoring system was meticulously set up with strategically positioned sensors at various river locations, ensuring data collection at regular 5-min intervals. This data was then transmitted to a cloud-based web portal, facilitating storage and analysis. To assess water quality, we introduced a novel hybrid approach, combining Convolutional Neural Networks (CNN) and Long Short-Term Memory (LSTM) networks. The proposed CNN-LSTM model achieved a validation accuracy of 98.40%, surpassing the performance of other state-of-the-art methods. Notably, the practical application of this system includes real-time alerts, promptly notifying stakeholders when water quality parameters exceed predefined thresholds. This feature aids in making informed decisions in water resource management. The study's contributions lie in its effective river water quality monitoring system, which encompassing various parameters, and its potential to positively impact environmental conservation efforts by providing a valuable tool for informed decision-making and timely interventions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The present disrupted scenario of the world calls for urgent attention to the need for renewable resources as an energy source for harnessing and feeding uninterrupted power supply to mankind. Amidst this, Photocatalysis (PC) and Photoelectrocatalysis (PEC) are some of the most budding methods of exploiting solar energy. LaFeO 3 -based systems are eligible for PC/PEC Hydrogen (H 2 ) generation, incorporating the process of water splitting, etc. It would be fair to mention that the above methods can mimic the natural process of photosynthesis. This review comprises an encyclopedia of recent advancements in LaFeO 3 and modified systems towards sustainable Photocatalytic and Photoelectrocatalytic Hydrogen Evolution Reactions (HER). Besides the challenges, the review presents a clear and brief idea for the scientific research community on paving the future in upscaling and industrializing the LaFeO 3 -mediated green fuel (H2) generation to meet global energy needs., Competing Interests: Declaration of competing interest There is no conflict of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The use of submucosal injection is crucial for satisfactory submucosal elevation in the early resection of flat polyps originating from the gastrointestinal tract (GIT). Injectable hydrogels derived from natural polypeptides are attractive candidates due to their excellent biocompatibility and easy gelation properties. However, most of the reported hydrogels are not the class of catheter delivery materials due to quick gelation, high inherent viscosity, and injection clogging. This study presents a novel injectable shear-thinning hydrogel platform of small molecules (nonanal) modified gelatin polymer, which offers a promising submucosal injection for effective removal of polyps from GIT. Physicochemical characterizations of hydrogel demonstrate the suitable features as an effective submucosal injection, including shear thinning property, self-assembly, methylene blue dye encapsulation, flow behavior, stability, syringeability (18 G, 21 G, and 24 G needles) and fibrous morphology. Ex vivo investigations of developed submucosal formulation on goat intestines demonstrate the enhanced visibility of cushions and the ability to produce stable, long-lasting cushions of about 8.07 mm up to ∼60 min of submucosal injection. The rapid blood clotting behavior of hydrogel was observed in about 120 s without compromising hemocompatibility with the hemolysis of about 3.77 % only. In vitro biocompatibility of the hydrogel was also verified using the HepG2 and nHDF cells. In vivo study depicts desirable biocompatibility, a non-toxic organ profile, and optimal cushion height in mice models. Studies established the foundation of novel submucosal fluid to improve the therapeutic outcomes of early resection for gastrointestinal polyps., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Diagnostic ultrasonography has evolved to become an indispensable imaging tool that permits non-invasive evaluation of the whole body. In this narrative review, we present a historical timeline of the invention, development, and evolution of diagnostic medical ultrasound. It includes interesting fun facts that may help the reader identify with many of the incredible researchers in this field. This review is a tribute to the researchers who contributed to this amazing invention., (© 2024 The Author(s). Journal of Clinical Ultrasound published by Wiley Periodicals LLC.)

Objective: There is increasing recognition that health systems need to measure and improve the value of patient care by measuring outcomes. Chronic pelvic pain secondary to pelvic venous insufficiency can have a significant impact on the quality of life (QOL) of women affected. Despite growing recognition, pelvic venous disorders (PeVDs), an important cause of chronic pelvic pain, remain underdiagnosed. Developing a core outcome set (COS) for benchmarking care delivery enhances the standardization of care. However, there is no consensus regarding a standardized minimum set of outcomes for PeVD. We aimed to generate a list of outcomes reported in previous PeVD treatment studies to lay the foundation for developing a COS for PeVD., Methods: This scoping review was undertaken according to the PRISMA-ScR guidelines. Initially, screening, full-text review and extraction was conducted on studies published between 2018 and 2023. Subsequently, the search was expanded using 1-year intervals, until, over a 1-year interval, no new outcomes were recorded. Closely related outcomes were classified into domains, and domains into three core areas: disease-specific, treatment-related, and QOL-related outcomes., Results: Of the 1579 records identified, 51 publications were included. From these studies, 108 different outcomes were identified. The median number of outcomes per study was 8 (interquartile range, 6-13). Closely related outcomes were organized into 42 outcome domains, which were then categorized into 3 core outcome areas; 47.6% (20/42) were disease specific, 35.7% (15/42) treatment related, and 16.7% (7/42) were QOL related. Of the 51 included studies, disease-specific outcomes were identified in 96.1% of the studies (49/51), treatment-related outcomes in 94.1% (48/51), and QOL outcomes in only 13.7% (7/51)., Conclusions: There was significant heterogeneity in outcomes reported in PeVD studies. Most PeVD treatment studies evaluated disease-specific and treatment-related outcomes of PeVD, but few reported outcomes that measured the impact on QOL. These findings will inform the next steps in developing a COS for PeVD., Competing Interests: Disclosures None., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Objective: National efforts have highlighted the need for pediatric emergency readiness across all settings where children receive care. Outpatient offices and urgent care centers are frequent starting points for acutely injured and ill children, emphasizing the need to maintain pediatric readiness in these settings. We aimed to characterize emergency medical services (EMS) utilization from outpatient offices and urgent care centers to better understand pediatric readiness needs., Methods: We performed a retrospective cross-sectional analysis of EMS encounters using the National Emergency Medical Services Information System, a nationally representative EMS registry (2019-2022). We included four years of EMS encounters of children (<18 years old) that originated from an outpatient office or urgent care center. We described characteristics, including patient demographics, prehospital clinician impression, therapies, and procedures performed., Results: Of 179,854,336 EMS encounters during the study period, 164,387 pediatric encounters originated at an outpatient setting. Most EMS encounters originated from outpatient offices. Evening and weekend EMS encounters more frequently originated from urgent care centers. The most common impressions were respiratory distress (n = 60,716), systemic illness (n = 23,583), and psychiatric/behavioral health (n = 13,273). Ninety-four percent of EMS encounters resulted in transportation to a hospital., Conclusions: EMS encounters from outpatient settings most commonly originate from outpatient offices, relative to urgent care settings, where pediatric emergency readiness may be limited. It is important that outpatient settings and providers are ready for varied emergencies, including those occurring for a behavioral health concern, and that readiness guidelines are updated to address these needs., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Academic Pediatric Association. Published by Elsevier Inc. All rights reserved.)

Background: Pelvic venous disorders (PeVD) encompass a variety of conditions linked to chronic pelvic pain in women. However, PeVD remain underdiagnosed due to the absence of universally accepted diagnostic criteria. The complexity of PeVD classifications across specialties leads to delays in treatment. This scoping review aims to fill a gap in PeVD diagnosis and management by identifying all existing scoring or grading systems to lay the foundation for standardized clinical scoring tools for PeVD., Methods: This scoping review was undertaken according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for Scoping reviews. Online databases were searched up to April 2023. Studies implementing a scoring or grading system for patients with confirmed or suspected PeVD were included. Scores or grading systems were classified into four main categories based on their use in the study: screening, diagnosis, measure of disease severity, and measure of response to treatment., Results: Of the 2976 unique records identified, 82 were reviewed in full, and 20 were included in this study. The publication dates ranged from 1984 to 2023 (median, 2018; interquartile range, 2003-2022). A total of 21 scores and/or grading systems were identified. Of these 21 scores, 10 (47.6%) were clinical scores, and 10 (47.6%) were scores based on radiological findings; one study included a score that used both clinical and radiological findings. The identified scores were used in various settings. Of the 21 scores, 2 (9.52%) were used for screening in a tertiary care setting; 3 (14.3%) were used to establish the PeVD diagnosis; 8 (38.1%) were used to assess disease severity; and 8 (38.1%) were used as measures of response to treatment. Of the eight scores assessing disease severity, four (50.0%) assessed the degree of dilatation of pelvic veins and four (50%) assessed the severity of reflux. Only three of the scores were validated., Conclusions: This scoping review identified a range of scoring and grading systems for PeVD. We note a lack of a validated scoring system, both clinical and radiological, for screening and assessment of disease severity. This is an important first step in developing validated disease-specific scoring systems for patient screening, appropriate referral, assessment of symptom severity, and assessment of the response to treatment., Competing Interests: Disclosures None., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Importance: Cardiac amyloid infiltration is the key determinant of survival in systemic light-chain (AL) amyloidosis. Current guidelines recommend early switching therapy in patients with a nonoptimal or suboptimal response regardless of the extent of cardiac amyloid infiltration., Objective: To assess the differences between serum biomarkers, echocardiography, and cardiovascular magnetic resonance (CMR) with extracellular volume (ECV) mapping in characterizing cardiac amyloid, the independent prognostic role of these approaches, and the role of ECV mapping to guide treatment strategies., Design, Setting, and Participants: Consecutive patients newly diagnosed with systemic AL amyloidosis (2015-2021) underwent echocardiography, cardiac biomarkers, and CMR with ECV mapping at diagnosis. Data were analyzed from January to June 2024., Main Outcomes and Measures: The primary outcomes of the study were all-cause mortality and hematological response as defined according to validated criteria: no response (NR), partial response (PR), very good partial response (VGPR), and complete response (CR). Secondary outcomes were the depth and speed of hematological response and overall survival according to ECV., Results: Of 560 patients with AL amyloidosis, the median (IQR) age was 68 years (59-74 years); 346 patients were male (61.8%) and 214 female (38.2%). Over a median (IQR) 40.5 months 9-58 months), ECV was independently associated with mortality. In the landmark analysis at 1 month, long-term survival was independent of the achieved hematological response in ECV less than 0.30% and ECV of 0.31% to 0.40%, while it was dependent on the depth of the hematological response in ECV greater than 0.40%. In the landmark analysis at 6 months, survival was independent of the achieved hematological response in ECV less than 0.30% and dependent on achieving at least PR in ECV of 0.31% to 0.40%. Survival was dependent on achieving CR in ECV of 0.41% to 0.50% and ECV greater than 0.50%. Achieving a deep hematological response at 1 month was associated with better survival compared with 6 months in patients with ECV greater than 0.40% but not with ECV less than 0.40%., Conclusions and Relevance: This study found that ECV mapping, in systemic AL amyloidosis, is an independent predictor of prognosis, can help define the hematological response associated with better long-term outcomes for each patient and potentially inform treatment strategies.

Land managers, researchers, and regulators increasingly utilize environmental DNA (eDNA) techniques to monitor species richness, presence, and absence. In order to properly develop a biological assay for eDNA metabarcoding or quantitative PCR, scientists must be able to find not only reference sequences (previously identified sequences in a genomics database) that match their target taxa but also reference sequences that match non-target taxa. Determining which taxa have publicly available sequences in a time-efficient and accurate manner currently requires computational skills to search, manipulate, and parse multiple unconnected DNA sequence databases. Our team iteratively designed a Graphic User Interface (GUI) Shiny application called the Reference Sequence Browser (RSB) that provides users efficient and intuitive access to multiple genetic databases regardless of computer programming expertise. The application returns the number of publicly accessible barcode markers per organism in the NCBI Nucleotide, BOLD, or CALeDNA CRUX Metabarcoding Reference Databases. Depending on the database, we offer various search filters such as min and max sequence length or country of origin. Users can then download the FASTA/GenBank files from the RSB web tool, view statistics about the data, and explore results to determine details about the availability or absence of reference sequences., Competing Interests: We have no Competing Interests to report., (Copyright: © 2024 Ramesh et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Necroptosis is a regulated form of cell death that has been observed in Alzheimer's disease (AD) along with the classical pathological hallmark lesions of amyloid plaques and Tau neurofibrillary tangles. To understand the neurodegenerative process in AD, we studied the role of necroptosis in mouse models and primary mouse neurons. Using immunohistochemistry, we demonstrated activated necroptosis-related proteins in transgenic mice developing Tau pathology and in primary neurons from amyloid precursor protein (APP)-Tau double transgenic mice treated with phosphorylated Tau seeds derived from a patient with AD but not in APP transgenic mice that only exhibited β-amyloid deposits. Necroptosis proteins in granulovacuolar degeneration (GVD) bodies were associated with neuronal loss in mouse brain regions also known to be vulnerable to GVD in the human AD brain. Necroptosis inhibitors lowered the percentage of neurons showing GVD and reduced neuronal loss, both in transgenic mice and in primary mouse neurons. This suggests that a GVD-associated form of necroptosis that we refer to as "GVD-necroptosis" may represent a delayed form of necroptosis in AD. We propose that inhibition of necroptosis could rescue this type of neuronal death in AD.

Since the recent discovery of polar topologies, a recurrent question has been how to remotely tune them. Many efforts have focused on the pumping of polar optical phonons from optical methods, but with limited success, as only switching between specific phases has been achieved so far. Additionally, the correlation between optical pulse characteristics and the resulting phase is poorly understood. Here, we propose an alternative approach and demonstrate the deterministic and dynamical tailoring of polar topologies using acoustic phonon excitations. Our second-principles simulations reveal that by pumping specific longitudinal and transverse acoustic phonons, various topological textures can be induced in materials like BaTiO 3 or PbTiO 3 . This method leverages the strong coupling between polarization and strain in these materials, enabling predictable and dynamic control of polar patterns. Our findings open up an alternative possibility for the manipulation of polar textures, suggesting a promising research direction.

Cardiovascular disease (CVD) remains a global economic burden even in the 21st century with 85% of deaths resulting from heart attacks. Despite efforts in reducing the risk factors, and enhancing pharmacotherapeutic strategies, challenges persist in early identification of disease progression and functional recovery of damaged hearts. Targeting mitochondrial dysfunction, a key player in the pathogenesis of CVD has been less successful due to its role in other coexisting diseases. Additionally, it is the only organelle with an agathokakological function that is a remedy and a poison for the cell. In this review, we describe the origins of cardiac mitochondria and the role of heteroplasmy and mitochondrial subpopulations namely the interfibrillar, subsarcolemmal, perinuclear, and intranuclear mitochondria in maintaining cardiac function and in disease-associated remodeling. The cumulative evidence of mitochondrial retrograde communication with the nucleus is addressed, highlighting the need to study the genotype-phenotype relationships of specific organelle functions with CVD by using approaches like genome-wide association study (GWAS). Finally, we discuss the practicality of computational methods combined with single-cell sequencing technologies to address the challenges of genetic screening in the identification of heteroplasmy and contributory genes towards CVD., (© 2024. The Author(s).)

Planar cell polarity (PCP) - tissue-scale alignment of the direction of asymmetric localization of proteins at the cell-cell interface - is essential for embryonic development and physiological functions. Abnormalities in PCP can result in developmental imperfections, including neural tube closure defects and misaligned hair follicles. Decoding the mechanisms responsible for PCP establishment and maintenance remains a fundamental open question. While the roles of various molecules - broadly classified into "global" and "local" modules - have been well-studied, their necessity and sufficiency in explaining PCP and connecting their perturbations to experimentally observed patterns have not been examined. Here, we develop a minimal model that captures the proposed features of PCP establishment - a global tissue-level gradient and local asymmetric distribution of protein complexes. The proposed model suggests that while polarity can emerge without a gradient, the gradient not only acts as a global cue but also increases the robustness of PCP against stochastic perturbations. We also recapitulated and quantified the experimentally observed features of swirling patterns and domineering non-autonomy, using only three free model parameters - the rate of protein binding to membrane, the concentration of PCP proteins, and the gradient steepness. We explain how self-stabilizing asymmetric protein localizations in the presence of tissue-level gradient can lead to robust PCP patterns and reveal minimal design principles for a polarized system., Competing Interests: DS, SR, MJ, MR The authors declare that no competing interests exist., (© 2024, Singh et al.)

Introduction: Mesenchymal stem cell-derived extracellular vesicles (MSC EVs) hold significant promise for regenerative medicine. Lyophilization of EVs significantly enhances their translational potential. While, lyophilized EVs have been studied from a morphological perspective, the functional stability of these EVs and their cargo following lyophilization need to be mechanistically investigated., Methods: In this study, we investigated the functional and mechanistic bioactivity of fresh versus lyophilized MSC EVs, specifically focusing on functionally engineered osteoinductive EVs developed in our laboratory. We utilized dimethyl sulfoxide (DMSO) as a cryoprotectant and conducted pathway-specific in vitro and in vivo experiments to assess the stability and functionality of the EVs., Results: Our findings show that using DMSO as a cryoprotectant before lyophilization preserves the functional stability of engineered MSC EVs. In vitro experiments demonstrated that the endocytosis, cargo integrity, and pathway-specific activity of lyophilized EVs were maintained when DMSO was used as the cryoprotectant. Additionally, in vivo bone regeneration studies revealed that the functionality of cryoprotected lyophilized EVs was comparable to that of freshly isolated EVs., Discussion: These results provide a foundation for evaluating the functionality of lyophilized EVs and exploring the use of DMSO and other cryoprotectants in EV-based therapies. Understanding the functionality of lyophilized naïve and engineered EVs from a mechanistic perspective may enhance validation approaches for tissue regeneration strategies., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Huang, Kang, Debnath, Leung, Raghavan, Lu, Cooper, Gajendrareddy and Ravindran.)

Deep learning Neural Networks (NN) have been developed in the field of molecular modeling for the purpose of circumventing the high computational cost of quantum-mechanical calculations while rivaling their accuracies. Although these networks have found great success, they generally lack the ability to accurately describe long-range interactions, which makes them unusable for extended molecular systems. Herein, we provide a method for partially retraining the deep learning general-use neural network ANI, in which the long-range interactions are represented via atomic electrostatic potentials. The electrostatic potentials, generated with polarizable effective fragment potentials (EFP), are used as an additional input feature for the network. This new ANI/EFP network can predict solute-solvent interaction energies on a trained data set with a kcal/mol accuracy. It also shows promise in predicting the interaction energies of a solute in solvent environments that have not been included in a training data set. The proposed protocol can be taken as an example and further developed, leading to highly accurate and transferable neural network potentials capable of handling long-range interactions and extended molecular systems.

Objective This study aims to identify the risk factors associated with postoperative complications following standard percutaneous nephrolithotomy (PCNL) for better results with the emergence of mini-percutaneous nephrolithotomy (mini-PCNL)/retrograde intrarenal surgery (RIRS) in recent years. Methodology This retrospective study was conducted in the Department of Urology, Sri Ramachandra Institute of Higher Education and Research, tertiary health care center in Chennai, India, from January 2018 to December 2023. Records of demographic information, along with clinical presentations like any urinary tract infection (UTI), hematuria, or loin pain were recorded. Medical and surgical history noted. Characteristics such as stone burden, location, number of calyces involved, and partial or complete staghorn were noted. Patients had undergone standard PCNL, either prone or supine, as per the preference of the surgeon. Operative time, number of percutaneous puncture tracts, and other intraoperative factors were investigated. A modified Clavien-Dindo scoring system for PCNL was used to classify any complication that occurred during this time. All patients were assessed after one month for stone clearance status. Results IBM SPSS Statistics for Windows, version 29 (IBM Corp., Armonk, NY, USA) was used for statistical analysis. The logistic regression analysis results show that age >55 years, urinary tract infection (UTI), diabetes, CKD, stone burden, percutaneous access number >1, and Amplatz sheath size >30Fr are highly significant predictors of complications (p <0.05). A greater number of calyces involved significantly increases the odds of complications. The odds ratio is 2.307 with a significant p-value (0.045), indicating a significant association. Longer operative time substantially increases the odds of complications (odds ratio is 0.129 with p = 0.021). Conclusion Standard PCNL has a low overall complication rate and is still an excellent modality of treatment for large-burden kidney stones in the mini-PCNL/RIRS era. Preoperative optimization, reduction in percutaneous access tracts with smaller Amplatz sheath (24Fr), and shorter operative time will help to reduce the complications rate., Competing Interests: Human subjects: Consent was obtained or waived by all participants in this study. Animal subjects: All authors have confirmed that this study did not involve animal subjects or tissue. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work., (Copyright © 2024, Aher et al.)

H3K27M diffuse midline gliomas (DMG), including diffuse intrinsic pontine gliomas (DIPG), exhibit cellular heterogeneity comprising less-differentiated oligodendrocyte precursors (OPC)-like stem cells and more differentiated astrocyte (AC)-like cells. Here, we establish in vitro models that recapitulate DMG-OPC-like and AC-like phenotypes and perform transcriptomics, metabolomics, and bioenergetic profiling to identify metabolic programs in the different cellular states. We then define strategies to target metabolic vulnerabilities within specific tumor populations. We show that AC-like cells exhibit a mesenchymal phenotype and are sensitized to ferroptotic cell death. In contrast, OPC-like cells upregulate cholesterol biosynthesis, have diminished mitochondrial oxidative phosphorylation (OXPHOS), and are accordingly more sensitive to statins and OXPHOS inhibitors. Additionally, statins and OXPHOS inhibitors show efficacy and extend survival in preclinical orthotopic models established with stem-like H3K27M DMG cells. Together, this study demonstrates that cellular subtypes within DMGs harbor distinct metabolic vulnerabilities that can be uniquely and selectively targeted for therapeutic gain., (© 2024. The Author(s).)

Study Objective: To evaluate for increases in the use and costs of respiratory viral testing in pediatric emergency departments (EDs) because of the COVID-19 pandemic., Methods: We performed a cross-sectional study using the pediatric health information system. Eligible subjects were children (90 days to 18 years) who were discharged from a pediatric ED and included in the pediatric health information system from October 2016 through March 2024. To evaluate for changes in the frequency and costs of respiratory viral testing, we performed an interrupted time series analysis across 3 study periods: prepandemic (October 1, 2016 to March 14, 2020), early pandemic (March 15, 2020 to December 31, 2023), and late pandemic (January 1, 2023 to March 31, 2024)., Results: We included 15,261,939 encounters from 34 pediatric EDs over the 90-month study period. At least 1 viral respiratory test was performed for 460,826 of 7,311,177 prepandemic encounters (6.3%), 1,240,807 of 5,100,796 early pandemic encounters (24.3%), and 545,696 of 2,849,966 late pandemic encounters (19.1%). There was a positive prepandemic slope in viral testing (0.17% encounters/month; 95% CI 0.17 to 0.18). The early pandemic was associated with a shift change of 4.98% (95% CI 4.90 to 5.07) and a positive slope (0.54% encounters/month; 95% CI 0.54 to 0.55). The late pandemic period was associated with a negative shift (-17.80%; 95% CI -17.90 to -17.70) and a positive slope (0.42% encounters/month; 95% CI 0.41 to 0.42). The slope in testing costs increased from $5,000/month (95% CI $4,200 to $5,700) to $33,000/month (95% CI $32,000 to $34,000) during the early pandemic., Conclusion: Respiratory testing and associated costs increased during the COVID-19 pandemic and were sustained despite decreasing incidence of disease. These findings highlight a need for further efforts to clarify indications for viral testing in the ED and efforts to reduce low-value testing., (Copyright © 2024 American College of Emergency Physicians. Published by Elsevier Inc. All rights reserved.)

Context: Tumor volume doubling time (TVDT) is emerging as a useful tool in predicting oncologic outcomes. There is limited data on the prognostic role of TVDT in metastatic medullary thyroid cancer (MTC)., Purpose: The goal of this study was to assess the value of TVDT in predicting disease-specific survival (DSS) in patients with hereditary and sporadic MTC., Methods: This was an Institutional Review Board-approved cohort study including patients with metastatic MTC having at least 3 consecutive imaging studies. TVDT of up to the five largest lesions per organ was calculated using a standardized formula. The association between TVDT and DSS was analyzed using Kaplan-Meier survival curves. Cox proportional regression model was used to account for confounding factors., Results: The study sample consisted of 51 patients presenting with 286 metastatic lesions measured with 457 scans during the follow-up of 51 (IQR 25-102) months. Median age was 19 years (IQR 15-41), 53% female patients. Cumulative volumes of all metastatic lesions and proportion of patients with TVDT of <1 year were higher in patients with sporadic as compared with hereditary MTC (p<0.01). Factors independently associated with shorter DSS were TVDT of <1 year based on 3 initial and 3 last scans as well as lung, brain and prostate as the organs with the fastest growing tumor. TVDT based on 2-dimentional and 3-dimentional measurements showed strong correlation (r=0.94, p<0.05)., Conclusions: Three baseline and three most recent scans preceding follow-up visit enable calculation of TVDT and can be used as predictors of mortality from MTC., (Published by Oxford University Press on behalf of the Endocrine Society 2024.)

Systemic sclerosis is a connective tissue disorder characterized by excessive fibrosis that primarily affects women, and can present as a multisystem pathology. Roughly 4-22% of patients with systemic sclerosis develop cancer, which drastically worsens prognosis. However, the mechanisms underlying systemic sclerosis initiation, propagation, and cancer development are poorly understood. We hypothesize that the inflammation and immune response associated with systemic sclerosis can trigger DNA damage, leading to elevated somatic mutagenesis, a hallmark of pre-cancerous tissues. To test our hypothesis, we culture clonal lineages of fibroblasts from the lung tissues of controls and systemic sclerosis patients and compare their mutation burdens and spectra. We find an overall increase in all major mutation types in systemic sclerosis samples compared to control lung samples, from small-scale events such as single base substitutions and insertions/deletions, to chromosome-level changes, including copy-number changes and structural variants. In the genomes of patients with systemic sclerosis, we find evidence of somatic hypermutation or kategis (typically only seen in cancer genomes), we identify mutation signatures closely resembling the error-prone translesion polymerase Polη activity, and observe an activation-induced deaminase-like mutation signature, which overlaps with genomic regions displaying kataegis., (© 2024. The Author(s).)

Spontaneous herniation of the temporomandibular joint into the external auditory canal is rare and generally results from inflammation or trauma but is rarely documented as a result of osteoradionecrosis. We report the novel surgical management of TMJ herniation by reconstructing the anterior EAC using conchal cartilage and a vascularized pedicled muscle flap in two patients. At follow-up, both had healing of the ear canal and TM without any evidence of joint herniation, with improved QOL and hearing. TMJ herniation due to an anterior EAC defect from osteoradionecrosis or trauma can be reconstructed with a cartilage graft and vascularized fascia flap. Laryngoscope, 2024., (© 2024 The American Laryngological, Rhinological and Otological Society, Inc.)

Time-resolved serial femtosecond crystallography (TR-SFX) of biological molecules captures the time-evolved dynamics of the residual motions across crystal structures, enabling the visualization of structural changes in response to chemical and physical stimuli to elucidate the relationship between the structure and function of the system under study. However, interpretations of residual motions can be complex to deconvolute because of various factors such as the system's size, temporal and spatial complexity, and allosteric behavior away from active sites. Relying solely on electron density map visualization can also pose a challenge in differentiating between useful and irrelevant data. In order to accurately identify residues and determine their respective contributions to the reaction dynamics, new tools are needed. We developed a new tool, ResiDEM, which employs a clustering-based approach to group difference electron densities and associate them with proximal residues. It can identify and rank residues with significant motions. Network representation can be used to delineate the interrelations between the residues in motion. With these features, ResiDEM helps to interpret residual motions in TR-SFX data, identify key residues, and elucidate their roles in dynamic processes.

Co-transcriptional assembly is an integral feature of the formation of RNA-protein complexes that mediate translation. For ribosome synthesis, prior studies have indicated that the strict order of transcription of rRNA domains may not be obligatory during bacterial ribosome biogenesis, since a series of circularly permuted rRNAs are viable. In this work, we report the structural insights into assembly of the bacterial ribosome large subunit (LSU) based on cryo-EM density maps of intermediates that accumulate during in vitro ribosome synthesis using a set of circularly permuted (CiPer) rRNAs. The observed ensemble of 23 resolved ribosome large subunit intermediates reveals conserved assembly routes with an underlying hierarchy among cooperative assembly blocks. There are intricate interdependencies for the formation of key structural rRNA helices revealed from the circular permutation of rRNA. While the order of domain synthesis is not obligatory, the order of domain association does appear to proceed with a particular order, likely due to the strong evolutionary pressure on efficient ribosome synthesis. This work reinforces the robustness of the known assembly hierarchy of the bacterial large ribosomal subunit and offers a coherent view of how efficient assembly of CiPer rRNAs can be understood in that context., (© The Author(s) 2024. Published by Oxford University Press on behalf of Nucleic Acids Research.)

This cross-sectional analysis of 86,111 visits for sickle cell disease and vaso-occlusive episodes (VOE) in U.S. pediatric emergency departments between 2013 and 2023 shows increased use of NSAIDs, ketamine, and acetaminophen, with unchanged opioid use. Hospitals with a higher volume of VOE visits more frequently administered opioids., Background: Vaso-occlusive episodes (VOEs) are a hallmark of sickle cell disease (SCD), leading to frequent emergency department (ED) visits. Effective pain management is crucial, with guidelines recommending routine use of non-steroidal anti-inflammatory drugs (NSAIDs) with opioids, and emerging evidence supporting ketamine use. However, these recommendations are based on low-certainty evidence, and the impact of these guidelines on analgesia use over time remains unclear., Objective: This study aimed to analyze trends in analgesia use over an 11-year period in pediatric SCD patients presenting to U.S. EDs with VOE and assess variations in treatment across hospitals., Methods: A cross-sectional study was conducted using data from the Pediatric Health Information System covering 34 U.S. children's hospitals from January 1, 2013, to December 31, 2023. The primary outcomes were the proportions of visits where opioids, NSAIDs, acetaminophen, and/or ketamine were administered on the first calendar day of the initial visit. Secondary outcomes included the co-administration of NSAIDs with opioids. Logistic and linear regression models were used to assess trends and hospital-level variations., Results: A total of 86,111 ED visits for VOE were analyzed. Opioids were administered in 82 % of encounters, NSAIDs in 72 %, acetaminophen in 17 %, and ketamine in 1 %. Co-administration of NSAIDs with opioids occurred in 59 % of the visits. Among discharged patients, there was a positive trend for NSAID use (slope: 1.68 %/year, 95 % CI: 0.91 %, 2.45 %) and NSAID-opioid co-administration (slope: 1.03 %/year, 95 % CI: 0.37 %, 1.69 %) over time. Acetaminophen use also increased over the study period (slope: 0.99 %/year, 95 % CI: 0.80 %, 1.17 %). In hospitalized patients, there was a significant upward trend for acetaminophen (slope: 1.29 %/year, 95 % CI: 0.69 %, 1.89 %) and ketamine (slope: 0.36 %/year, 95 % CI: 0.27 %, 0.45 %), while opioid use remained unchanged. Significant hospital-level variations were observed, with larger hospitals more likely to administer opioids but less likely to co-administer NSAIDs with opioids compared to medium-volume hospitals., Conclusion: Over the past decade, the use of NSAIDs, acetaminophen, and ketamine has increased in the management of VOE in pediatric SCD patients, while opioid use remains consistent. The co-administration of NSAIDs and opioids has also increased, reflecting guideline adherence. Variations in analgesia practices across hospitals underscore the need for standardizing pain management strategies in this population., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Inc.)

We discover an instability mechanism in suspensions of self-propelled particles that does not involve active stress. Instead, it is driven by a subtle interplay of inertia, swimmer motility, and concentration fluctuations, through a crucial time lag between the velocity and the concentration field. The resulting time-persistent state seen in our high-resolution numerical simulations consists of self-sustained waves of concentration and orientation, transiting from regular oscillations to wave turbulence. We analyze the statistical features of this active turbulence, including an intriguing connection to the Batchelor spectrum of passive scalars.

Objective: Measures of neighborhood disadvantage demonstrate correlations to health outcomes in children. We compared differing indices of neighborhood disadvantage with emergency medical services (EMS) interventions in children., Methods: We performed a retrospective study of EMS encounters for children (<18 years) from approximately 2000 US EMS agencies between 2021 and 2022. Our exposures were the Child Opportunity Index (COI; v2.0), 2021 Area Deprivation Index (ADI), and 2018 Social Vulnerability Index (SVI). We evaluated the agreement in how children were classified with each index using the intraclass correlation coefficient. We used logistic regression to evaluate the association of each index with transport status, presence of cardiac arrest, and condition-specific interventions and assessments., Results: We included 738,892 encounters. The correlation between the indices indicated good agreement (intraclass correlation coefficient=0.75). There was overlap in relationships between the COI, ADI, and SVI for each of the study outcomes, both when visualized as a splined predictor and when using representative odds ratios (OR) comparing the third quartile of each index to the lower quartile (most disadvantaged). For example, the OR of non-transport was 1.12 (95% confidence interval [CI]: 1.10-1.14) for COI, 1.18 (95% CI: 1.16-1.20) for ADI, and 1.22 (95% CI: 1.20-1.23) for SVI., Conclusion: The COI, ADI, and SVI had good correlation and demonstrated similar effect size estimates for a variety of clinical outcomes. While investigators should consider potential causal pathways for outcomes when selecting an index for neighborhood disadvantage, the relative strength of association between each index and all outcomes was similar., Competing Interests: Declaration of Competing Interest The authors have indicated they have no potential conflicts of interest to disclose., (Copyright © 2024 Academic Pediatric Association. Published by Elsevier Inc. All rights reserved.)

Our knowledge of the contribution of genetic interactions ( epistasis ) to variation in human complex traits remains limited, partly due to the lack of efficient, powerful, and interpretable algorithms to detect interactions. Recently proposed approaches for set-based association tests show promise in improving the power to detect epistasis by examining the aggregated effects of multiple variants. Nevertheless, these methods either do not scale to large Biobank data sets or lack interpretability. We propose QuadKAST, a scalable algorithm focused on testing pairwise interaction effects ( quadratic effects ) within small to medium-sized sets of genetic variants (window size ≤100) on a trait and provide quantified interpretation of these effects. Comprehensive simulations show that QuadKAST is well-calibrated. Additionally, QuadKAST is highly sensitive in detecting loci with epistatic signals and accurate in its estimation of quadratic effects. We applied QuadKAST to 52 quantitative phenotypes measured in ≈300,000 unrelated white British individuals in the UK Biobank to test for quadratic effects within each of 9515 protein-coding genes. We detect 32 trait-gene pairs across 17 traits and 29 genes that demonstrate statistically significant signals of quadratic effects (accounting for the number of genes and traits tested). Across these trait-gene pairs, the proportion of trait variance explained by quadratic effects is comparable to additive effects, with five pairs having a ratio >1. Our method enables the detailed investigation of epistasis on a large scale, offering new insights into its role and importance., (© 2024 Fu et al.; Published by Cold Spring Harbor Laboratory Press.)

SNP heritability, the proportion of phenotypic variation explained by genotyped SNPs, is an important parameter in understanding the genetic architecture underlying various diseases and traits. Methods that aim to estimate SNP heritability from individual genotype and phenotype data are limited by their ability to scale to Biobank-scale data sets and by the restrictions in access to individual-level data. These limitations have motivated the development of methods that only require summary statistics. Although the availability of publicly accessible summary statistics makes them widely applicable, these methods lack the accuracy of methods that utilize individual genotypes. Here we present a SUMmary-statistics-based Randomized Haseman-Elston regression (SUM-RHE), a method that can estimate the SNP heritability of complex phenotypes with accuracies comparable to approaches that require individual genotypes, while exclusively relying on summary statistics. SUM-RHE employs Genome-Wide Association Study (GWAS) summary statistics and statistics obtained on a reference population, which can be efficiently estimated and readily shared for public use. Our results demonstrate that SUM-RHE obtains estimates of SNP heritability that are substantially more accurate compared with other summary statistic methods and on par with methods that rely on individual-level data., (© 2024 Jeong et al.; Published by Cold Spring Harbor Laboratory Press.)

Background: Initial oral antibiotics may be as effective as intravenous (IV) antibiotics for children hospitalized with community-acquired pneumonia (CAP), but further data are needed., Objective: We evaluated for associations of initial antibiotic route (IV vs. oral) with length of stay (LOS) and secondary outcomes for children hospitalized with CAP., Methods: This multicenter, retrospective cohort study included children with CAP who were hospitalized for >48 h, had chest radiographs, and received antibiotics at four children's hospitals between 2014 and 2020. Data were obtained from the Pediatric Health Information System and manual chart review. The exposure was initial antibiotic route (i.e., first antibiotic given intravenously or orally). We performed multivariable regression modeling using inverse probability treatment weights from propensity scores. Outcomes included LOS, oxygen duration, cost, care escalation, and readmission or emergency department revisit., Results: Of 1147 included children, 37% received initial oral antibiotics. Within the propensity balanced sample, LOS was 73.5 h (IQR 61.0, 99.5) and 78.7 (61.0, 118.0) for patients with initial oral and IV antibiotics, respectively. Children receiving initial oral antibiotics had an 8% reduction in LOS (OR 0.92 [95% CI: 0.87, 0.94]) and 14% reduction in cost (OR 0.86 [95% CI 0.79, 0.94]) versus those receiving initial IV antibiotics. There were no differences in other outcomes., Conclusions: Children with CAP receiving initial oral antibiotics had reduced LOS and hospital cost without differences in escalated care or return visits. Starting hospitalized children on oral antibiotics is likely a safe and effective alternative to IV treatment., (© 2024 Society of Hospital Medicine.)

Background: Root canal therapy in primary teeth is exceedingly complex due to the unique internal geometry of the pulp cavity., Aim: The purpose of this study was to evaluate variations in primary teeth root canal morphology and applicability of the Ahmed et al. (Int Endod J, 50, 2017, 761) alphanumeric system for classifying them., Design: A retrospective analysis of 2024 primary teeth from an existing CBCT database was performed for assessing variations in root canal morphology. Descriptive statistics and the chi-squared test were used for data analysis., Results: The maxillary canines exhibited splitting levels of 5.9% (right) and 6.6% (left), and merging levels of 1% (both sides); maxillary lateral incisors showed splitting levels of 2.6% (right) and 0.8% (left), with no merging observed; mandibular second molars had splitting levels of 1.5% (right) and 0.8% (left), with no merging observed; and mandibular lateral incisors exhibited a merging level of 0.9% (right), with no splitting observed., Conclusion: This study provides new insights into the variations in root canal morphology of primary teeth, particularly highlighting the variations found in primary canines. The Ahmed et al. (Int Endod J, 50, 2017, 761) classification system was found to be a useful tool for categorizing these variations., (© 2024 BSPD, IAPD and John Wiley & Sons Ltd.)

Introduction: Benign prostatic hyperplasia (BPH) is a common urological condition, particularly among middle-aged and elderly men. Trans urethral resection of prostate (TURP) has some drawbacks, particularly concerning ejaculatory function. Ejaculation preserving TURP (Ep-TURP) is one such technique that aims at preserving the tissues that are primarily responsible for antegrade ejaculation., Methods and Material: In this prospective study, patients with bothersome LUTS were randomized into Ep-TURP and standard TURP groups. Supramontal tissue was preserved in Ep-TURP while standard TURP group had resection of the entire prostatic tissue up to the verumontanum. The outcomes with regard to IPSS, Qmax, IIEF(Q9), Ejaculation and PVR were studied. Ejaculation projection score was separately calculated and assessed., Results: About 60 patients were studied, with 30 in each group. At 6 months' follow-up, complication rates were the same in both groups. The IPSS, Qmax, IIEF were similar and comparable. There was no change in erection in all patients (P = 0.559). The ejaculation was well preserved in Ep-TURP (p<0.001). Ejaculation projection score was maintained at 3.77 in the Ep-TURP group while in the standard group it was 0 (P<0.001)., Conclusions: Ep-TURP is a safe and a cost-effective method of preserving ejaculation in patients undergoing TURP. This procedure adds a better quality of life that greatly aids in enhancing the improving his psychological outlook towards life. Ep-TURP has now emerged as the standard procedure of choice in sexually active young males needing TURP, as the degree of ejaculation is largely well preserved in patients undergoing this procedure., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Purpose: Clinicians are often uncertain about their prognostic estimates, which may impede prognostic communication and clinical decision-making. We assessed the impact of a web-based prognostic calculator on physicians' prognostic confidence., Methods: In this prospective study, palliative care physicians estimated the prognosis of patients with advanced cancer in an outpatient clinic using the temporal, surprise, and probabilistic approaches for 6 m, 3 m, 2 m, 1 m, 2 w, 1 w, and 3 d. They then reviewed information from www.predictsurvival.com , which calculated survival estimates from seven validated prognostic scores, including the Palliative Prognostic Score, Palliative Prognostic Index, and Palliative Performance Status, and again provided their prognostic estimates after calculator use. The primary outcome was prognostic confidence in temporal CPS (0-10 numeric rating scale, 0 = not confident, 10 = most confident)., Results: Twenty palliative care physicians estimated prognoses for 217 patients. The mean (standard deviation) prognostic confidence significantly increased from 5.59 (1.68) before to 6.94 (1.39) after calculator use (p < 0.001). A significantly greater proportion of physicians reported feeling confident enough in their prognosis to share it with patients (44% vs. 74%, p < 0.001) and formulate care recommendations (80% vs. 94%, p < 0.001) after calculator use. Prognostic accuracy did not differ significantly before or after calculator use, ranging from 55-100%, 29-98%, and 48-100% for the temporal, surprise, and probabilistic approaches, respectively., Conclusion: This web-based prognostic calculator was associated with increased prognostic confidence and willingness to discuss prognosis. Further research is needed to examine how prognostic tools may augment prognostic discussions and clinical decision-making., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Erythema multiforme (EM) is an acute-onset, self-limiting inflammatory condition affecting the skin and mucous membranes. It exhibits a range of skin lesions, which is why it is referred to as "multiforme." Oral lesions, usually inflammatory and frequently marked by rapidly rupturing vesicles and bullae, often constitute a significant clinical feature. The exact etiology is obscure; however, it may be caused by multiple triggering factors. The most well-established connection is with a previous herpes simplex virus infection, while a smaller percentage of cases (<10%) are linked to drug use. Based on mucosal involvement, the condition is classified into two types: EM minor and EM major. Stevens-Johnson syndrome and toxic epidermal necrolysis (Lyell's disease) are now recognized as separate clinical conditions. EM usually exhibits a self-limiting course, with lesions typically improving within a few weeks. Avoiding triggers and using steroid therapy can be effective treatments. This paper documents a rare case of drug-induced oral EM featuring characteristic lip and intraoral lesions. The patient developed painful oral ulcers and difficulty in swallowing after taking azithromycin for a sore throat and fever. Clinical examination revealed multiple, hemorrhagic encrustations on the lips, along with ulcers on the right buccal mucosa and vesicular eruptions on the palate. The acute onset of multiple oral ulcers associated with a recent drug intake led to a diagnosis of drug-induced oral EM. Treatment included systemic prednisolone, topical triamcinolone paste, and supportive oral care. No recurrence was observed during the six-month follow-up., Competing Interests: Human subjects: Consent was obtained or waived by all participants in this study. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work., (Copyright © 2024, Sriram et al.)

Background: To explore provider perspectives about palliative care (PC) in head and neck cancer (HNC) care., Methods: A 25-question electronic survey was disseminated to the membership of the American Head and Neck Society (AHNS) from April 10, 2023, through June 13, 2023., Results: Respondents were most likely to refer to PC at symptomatic disease progression (52%) or terminal diagnosis (29%) rather than at initial diagnosis (17%). Participants less likely to refer to PC were less likely to refer to symptomatic progression (8% vs. 39%, p = 0.0006) or address advance directives (62% vs. 87%, p = 0.0406). Symptom burden questionnaires were used by only 29% of respondents. Discordance was identified between self-reported and actual access to local inpatient and outpatient PC services., Conclusions: Barriers to PC identified include a lack of established optimal timing of PC referral, a perceived lack of local access to PC, and a lack of uniform standardized assessment of symptom burden., (© 2024 Wiley Periodicals LLC.)