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The Generalized Anxiety Disorder 7-item scale (GAD-7) could provide currently lacking data on anxiety levels among adolescents and young adults in Sweden. However, psychometric assessments of the scale are scarce in this population. Additionally, the structural validity of the scale needs to be examined further, given conflicting findings in other populations. The aim of this study was to psychometrically evaluate the GAD-7 among 15-29-year-olds using Rasch analysis and confirmatory factor analysis in a Swedish, general population sample. In addition, an iterative process explored if revised versions of the scale could be identified that fit the Rasch model in this sample. A random, age- and sex-stratified sample of 15-29-year-olds from the Swedish population register was drawn, from which stratified groups were randomly invited to participate. Data (n = 590) were collected through a study-specific app. Rasch analyses included investigation of item fit, local independence, principal components analysis of standardized Rasch model residuals, response category functioning, measurement invariance, scale-to-sample targeting, and reliability. In addition, confirmatory factor analyses (CFA) were conducted and interpreted using dynamic, data-adaptive cutoffs. An exploratory, iterative process was undertaken aiming at scale refinement based on results for the GAD-7. No Rasch analyses provided support for the unidimensionality of the GAD-7, which also displayed issues with disordered response categories and measurement invariance. The CFA found substantial misfit for the one-factor model. The exploratory scale refinement resulted in two abbreviated scales warranting further investigation: Items 1-3 fit the Rasch model as a unidimensional scale and demonstrated acceptable to good reliability. Items 5-7 had good fit after merging the two highest response categories, although demonstrated poor reliability. No analyses provided support for the unidimensionality of the GAD-7 among 15-29-year-olds in Sweden. While these findings do not support the use of a single sum score, they require confirmation in independent samples. Future research should evaluate whether the abbreviated, candidate scale comprising items 1-3 could be useful to assess anxiety in this population, including examination of criterion validity and the potential benefits of adding a response category to enhance both reliability and scale-to-sample targeting.

Epstein-Barr virus (EBV) is highly prevalent worldwide and has been linked to different cancers and autoimmune disorders, including Multiple sclerosis (MS). Primary infection typically occurs during early childhood or adolescence and often goes undetected. Serological studies allow the detection of EBV-specific antibodies after infection, but venous blood collection is resource-intense and thus limits sample size. To overcome these challenges, we developed a high-throughput, semi-automated serology protocol targeting antibodies against EBV in capillary blood samples (Dried Blood Spots, DBS). Paired serum and DBS samples were obtained from 417 participants aged 18-25 years. We used Roche Elecsys® EBV nuclear antigen-1 EBNA IgG and Elecsys® EBV viral capsid antigen VCA IgG assays, established for serum, to detect antibodies against EBV anti-EBNA‑1 IgG and anti-VCA IgG in DBS. Cut-off Indices (COI) for the DBS assays were determined using a classification tree. Of the 416 valid paired serum samples, 78.6% (327/416) were anti-VCA-positive and 76.4% (318/416) anti-EBNA-positive. Discrepant results were observed in 3.6% (15/416) of participants. Based on combined assay results, 20.7% (86/416) of the samples were classified as EBV-negative. The serum COI values revealed that most of the negative cases clustered well below the cutoff, while positive cases spanned a broad range of higher values. Overall, 98.8% (326/330) of participants classified as positive based on serum testing (n = 330) were also classified as positive in DBS (n = 326), corresponding to the sensitivity of DBS relative to serum results. Likewise, 96.5% (83/86) of participants classified as serum-negative participants (n = 86) were also negative in DBS (n = 83), corresponding to the specificity of DBS relative to serum results. Among the 416 participants, only 4/330 (1.2%) were false negatives and 3/86 (3.5%) were false positives in DBS. The newly established assays were validated in a self-sampling cohort of 295 participants. We established DBS-specific cutoff values to detect antibodies against EBV in DBS, achieving high sensitivity (98.8%) and specificity (96.5%) relative to the corresponding venous blood assay. Furthermore, we derived a correction formula to convert semi-quantitative DBS values to serum-equivalents, enabling comparison with other studies and standardized datasets. The DBS-based approach allows sero-status assessment in a large population and thus e.g. simplifies the identification of suitable participants for clinical trials.

Neuroblastoma remains one of the most lethal pediatric solid tumors, and durable control of high-risk disease is hindered not only by inefficient tumor-selective drug delivery but also by a profoundly immunosuppressive tumor microenvironment. To address these dual barriers, we developed a chemoimmunotherapeutic vesicle-mimetic platform, DAS/CD40L-EM@DOX, by engineering CD40L-overexpressing HEK-293 T donor cells, generating extracellular vesicle mimetics through extrusion, decorating the membrane with a neuroblastoma-targeting DAS peptide, and post-loading doxorubicin. The resulting formulation preserved a nanoscale vesicular morphology, displayed CD40L on the membrane, and showed favorable particle size and zeta potential characteristics. In neuroblastoma cells, DAS decoration enhanced uptake in an α7 nicotinic acetylcholine receptor-associated manner and increased the cytotoxic and immunogenic effects of doxorubicin, as evidenced by augmented apoptosis, calreticulin exposure, and HMGB1 release. In macrophage assays, CD40L-containing vesicles shifted M2-like cells toward an M1-like phenotype, increasing CD86, TNF-α and IL-6 while decreasing CD206, IL-10 and TGF-β. Functionally, this repolarization enhanced tumor-cell phagocytosis and promoted CD8+ T-cell effector responses, including increased granzyme B, IFN-γ and IL-2 production. In tumor-bearing mice, DAS modification improved tumor accumulation and reduced off-target sequestration relative to unmodified vesicles. Therapeutically, DAS/CD40L-EM@DOX exerted the strongest inhibition of tumor growth, reduced tumor burden, prolonged survival, increased intratumoural M1-like macrophages and CD8+ T cells, decreased M2-like macrophages and regulatory T cells, and enhanced immunogenic cell-death markers in situ. Together, these findings support DAS/CD40L-EM@DOX as a dual-function vesicle-mimetic nanomedicine that couples neuroblastoma-targeted chemotherapy with macrophage reprogramming to remodel the tumor immune microenvironment and improve antitumour efficacy.

Posterior fossa decompression in the setting of Chiari malformation is a common neurosurgical procedure in adolescents; however, the effect of obesity on its surgical outcomes is understudied. This is partly due to the lower incidence of obesity in adolescents requiring higher statistical power. Correspondingly, we sought to interrogate a national surgical database to aggregate a significant cohort size to elucidate possible relationships between obesity and surgical outcomes in this niche. The American College of Surgeons National Surgical Quality Improvement Program-Pediatric (ACS NSQIP-Pediatric) database from 2015-2024 was interrogated against selection criteria targeting patients aged 10-18&#xa0;years old surgically treated for Chiari malformation type 1. Adolescent weight was defined by body mass index into categories according to the definitions by Centers for Disease Control and Prevention (CDC). Thirty-day surgical outcomes were modeled using a multivariate logistic regression controlling for covariates across a primary model, and then across various sensitivity models. A total of 4257 patients satisfied selection criteria with a median age of 14.4&#xa0;years, 65% female and 35% male. Median BMI was 22.7&#xa0;kg/m2, and per CDC definitions 6% were underweight, 51% were healthy weight, 18% were overweight, and 28% were obese (Classes I, II and III). Overall, median length of stay was 3&#xa0;days, and rates of 30-day reoperation (any), reoperation (related), unplanned readmission and major complications were 3.0%, 2.4%, 5.4%, and 3.5%, respectively. On analysis, categorical obesity (vs. healthy weight/underweight) was independently associated with the 30-day major complication composite (aOR 1.93, p&#x2009;<&#x2009;0.001) and unplanned readmission (aOR 1.47, p&#x2009;=&#x2009;0.012). Of the complications, surgical site infection (p&#x2009;<&#x2009;0.001) and peripheral nerve injury (p&#x2009;=&#x2009;0.011) were statistically more common in the obese group than others. As a continuous predictor, each&#x2009;+&#x2009;1 SD increase in BMI-for-age z-score was independently associated with higher adjusted odds of 30-day major complication (aOR 1.25, p&#x2009;=&#x2009;0.004) and unplanned readmission (aOR 1.13, p&#x2009;=&#x2009;0.038). Finally, linear trend tests across all six CDC extended categories were significant for 30-day reoperation (any) (p&#x2009;=&#x2009;0.031), reoperation (related) (p&#x2009;=&#x2009;0.005), and unplanned readmission (p&#x2009;=&#x2009;0.017). In adolescents, there is a clear postoperative profile that exists 30-days after posterior fossa decompression for Chiari malformation based on weight and obesity. This profile is defined by increased morbidity rather than mortality, and future studies are needed to determine how these findings can be effectively integrated into preoperative evaluation and prehabilitation workup for patients to optimize surgical outcomes further.

Adeno-associated virus (AAV) vectors are widely used for neuroscience research. Labeling AAV mRNA by in situ hybridization can track AAV transduction with high sensitivity and without the need for reporter genes. However, it is challenging to detect similar mRNA transcripts when multiple AAVs are injected into the same animal, or when a single AAV produces multiple distinct transcripts. To address these challenges, we developed methods for sensitive labeling of small barcodes on AAV mRNA at single-transcript resolution. These small RNA barcodes (ranging from 40 to 44 bases) can be easily added to any AAV vector genome. We provide detailed step-by-step protocols for sensitive and flexible labeling of barcoded AAV mRNA by BaseScope in situ hybridization. We validated these labeling methods in male and female mice, and in a female rhesus macaque. RNA barcoding can improve the reliability of multiplexed in vivo screening: by adding one barcode to half of the AAVs in a pooled library and a second barcode to the other half, barcode labeling can identify cells that receive multiple AAVs without the need for reporter proteins. This approach is also useful for intersectional experiments that combine multiple AAV vectors: multiple unique barcodes can track the spatial distribution of each AAV and can separately label transcripts from different AAV genome conformations, such as before or after Cre-Lox recombination. These new methods for sensitive multiplexed labeling of AAV mRNA will support greater rigor and reliability in neuroscience research.Significance Statement AAV vectors are commonly used in neuroscience research. Modern experimental designs are increasingly complex, often injecting two or more AAVs into different regions, or screening a library of many unique AAVs injected together into a single animal. In such complex experiments, it is challenging to track spatial distribution and rates of co-infection among multiple AAV vectors, especially if the AAVs do not carry reporter genes. To address these challenges, we developed small RNA barcodes and methods for highly sensitive labeling of barcoded AAV mRNA. These small barcodes can be easily added to any AAV genome without altering the experimental design, supporting the assessment of injection targeting and co-infection rates among multiple AAVs without the need for reporter genes.

Hematopoietic cell transplantation (HCT) is a curative immunotherapy for hematologic malignancies such as acute myeloid leukemia, primarily through graft-versus-tumor (GVT) effects mediated by donor T cells. However, up to 60% of recipients develop graft-versus-host disease (GVHD), with intestinal involvement being a major contributor to morbidity and mortality. While CD8 T cells drive GVT, CD4 T helper (Th) cells are largely the key mediators of intestinal GVHD. Although IFN-&#x3b3;-producing Th1 cells have been implicated in this process, recent evidence suggests a more complex role for Th cell subtypes. In our study, we identify the transcription factor GATA3, often associated with Th2 function, as a key regulator of Th cell proliferation, gut persistence, and cytokine production. Specifically, donor cell GATA3 was required for the early production of gut&#x2011;damaging granzyme A by intestinal Th cells and sustained granulocyte-macrophage colony-stimulating factor (GM&#x2011;CSF) production at later phases of the intestinal GVHD response. GATA3&#x2011;mediated maintenance of GM&#x2011;CSF expression during these later stages of disease was associated with increased intestinal eosinophil recruitment and activation, features that have been linked to GVHD severity in humans. Collectively, our findings demonstrate that GATA3&#x2011;expressing Th cells contribute to intestinal GVHD pathogenesis by sustaining GM-CSF-driven inflammatory circuits and promoting chronic eosinophilia. Targeting GATA3, or its downstream effector pathways, may therefore represent a novel therapeutic strategy to attenuate intestinal GVHD while preserving the overall efficacy of HCT.

Neisseria gonorrhoeae is a global public health threat, exacerbated by the rapid dissemination of ceftriaxone-resistant strains harboring penA allele 60.001. Current diagnostics are limited by long turnaround times, reliance on thermal cycling equipment, and poor suitability for resource-limited settings. We developed an isothermal recombinase polymerase amplification assay coupled with a lateral flow strip (RPA-LFS) for separate identification of N. gonorrhoeae and the ceftriaxone resistance-associated SNP C932T (A311V) in penA 60.001 and limited closely related alleles. Species detection employed an RPA-Nfo probe targeting the conserved porA pseudogene. Resistance detection employed an allele-specific amplification refractory mutation system RPA (ARMS-RPA) strategy, incorporating artificial mismatches to specifically discriminate the C932T SNP. Assay performance was evaluated using genomic DNA of a panel of gonococcal and non-gonococcal strains and validated directly on unprocessed clinical urine samples. The porA RPA-LFS assay demonstrated a detection limit of 30 fg of genomic DNA with no cross-reactivity against closely related Neisseria species or other bacterial pathogens. The mismatch penA 60.001 RPA-LFS assay achieved a detection limit of 300 fg and exhibited absolute analytical specificity, with no amplification observed from wild-type or other mosaic penA alleles. Validation using 50 clinical urine samples yielded 100% concordance (95% CI: 92.9-100%) with gonorrhoea identification by reference nucleic acid amplification tests, with 100% sensitivity (95% CI: 87.1-100%) and 100% specificity (95% CI: 85.7-100%), and 100% concordance (95% CI: 85.7-100%) with resistance allele detection, with 100% sensitivity (95% CI: 67.6-100%) and 100% specificity (95% CI: 79.6-100%). This integrated RPA-LFS platform provides rapid detection of gonococcal infections and ceftriaxone resistance without requiring a thermal cycler, delivering visual results within 20 minutes. The assay represents a promising point-of-care solution for timely diagnosis, targeted antibiotic therapy, and resistance surveillance in resource-limited settings, though further validation on diverse specimen types and integration of an internal amplification control are needed.

Colorectal serrated lesions are precursors to colorectal cancer, with a high recurrence risk after endoscopic resection. Identifying risk factors for recurrence and establishing a prediction model are crucial for optimizing surveillance strategies. A single-center retrospective study included 164 patients with colorectal serrated lesions who had endoscopic follow-up information after the surgery. These cases were divided into a polyp recurrence group (79 cases) and a non-recurrence group (85 cases). Demographic data, medical histories, laboratory findings, endoscopic characteristics of the lesions, and follow-up durations were therefore collected. Cox regression identified independent risk factors, which were incorporated into a nomogram. Model performance was evaluated using C-index, calibration curves, and decision curve analysis. Multivariate analysis identified non-alcoholic fatty liver disease (HR 1.813, 95%CI 1.080-3.045), atrophic gastritis (HR 1.733, 95%CI 1.087-2.760), history of gallbladder abnormalities (HR 1.791, 95%CI 1.107-2.898) and elevated total bile acid levels (HR 1.023, 95%CI 1.003-1.043) as independent predictors of recurrence. The nomogram demonstrated good discrimination (C-index: 0.703-0.820) and clinical applicability (decision curve analysis net benefit: 27-80% threshold). This study established a validated prediction model for colorectal serrated lesions recurrence, highlighting metabolism-related factors as key determinants. Targeting these factors and applying the nomogram may enhance risk stratification and postoperative surveillance, potentially reducing colorectal cancer progression. Limitations include the retrospective design, single-center data, and lacking further validations.

Colorectal cancer (CRC) remains a leading cause of cancer-related morbidity and mortality worldwide, driven by complex genetic, epigenetic, and environmental factors. Despite advances in conventional treatments such as surgery, chemotherapy, and radiotherapy, therapeutic resistance and disease recurrence continue to limit long-term outcomes. This review provides a comprehensive overview of emerging and targeted therapeutic strategies in CRC, with emphasis on their molecular basis and clinical relevance. The key pathways involved in CRC pathogenesis, including the adenoma-carcinoma sequence, microsatellite instability, and CpG island methylator phenotype, are discussed to highlight their roles in disease progression and therapeutic targeting. Recent advances in targeted therapies, particularly those directed against vascular endothelial growth factor and epidermal growth factor receptor, along with the expanding role of immunotherapy, are critically examined. In addition, novel approaches such as gene- and RNA-based therapies, microbiome modulation, nanotechnology-driven drug delivery systems, and precision medicine strategies based on multi-omic profiling are explored. Despite these developments, the challenges including tumor heterogeneity, therapeutic resistance, and limitations in drug delivery and biomarker identification remain significant. Future perspectives emphasize the integration of molecular profiling and innovative therapeutic platforms to enable more personalized and effective treatment strategies. Collectively, these advances highlight a shift toward precision oncology for improved management of colorectal cancer.

The present study comprehensively dissects the molecular landscape of elderly mantle cell lymphoma (MCL) patients enrolled in the phase II V-RBAC trial of the Fondazione Italiana Linfomi. Of the 140 patients enrolled in the trial, 132 had available gDNA extracted from lymph node biopsies or bone marrow aspirates and were included in the analysis. A CAPP-Seq assay targeting 146 genes relevant to MCL pathogenesis was employed to identify gene mutations and copy number variations. ATM was the most frequently mutated gene, detected in 55 patients (41.7%), followed by TP53 and KMT2D in 31 patients (23.5%). ATM deletion was observed in 32 patients (24%), while CDKN2A loss in 29 (22%). Beyond TP53 mutations, three other molecular lesions, including CDKN2A loss, CD36 mutations and single-hit ATM abnormalities (either mutation or deletion) were independently associated with progression-free survival after adjustment for high-risk trial-defining features, namely Ki-67 >30% and blastoid variant. Notably, patients harboring single-hit ATM alterations without any additional risk factors achieved durable long-term remission, while CD36 mutations were associated with adverse survival. Both findings represent previously unrecognized aberrations that in this cohort independently and inversely associated with survival. The four variables were integrated into a 4-factor molecular prognostic model internally validated using a bootstrapping approach, which identified four distinct patient subgroups with significantly different outcomes. These findings support the importance of i) molecular profiling in MCL, ii) risk-adapted trials like V-RBAC, and iii) the integration of other biological markers with TP53 mutations for a more precise risk assessment in MCL. (NCT03567876).

Despite the notable clinical advances brought about by programmed cell death ligand 1/programmed death 1 (PD-1/PD-L1) inhibitors in hepatocellular carcinoma (HCC) therapy, the tumor immunosuppressive microenvironment (TIME) exerts a restrictive effect on their overall therapeutic performance. To overcome the constraints imposed by TIME, this study developed an innovative targeted therapeutic strategy leveraging ultrasound-responsive nanobubbles (NBs) for the co-delivery of curcumin and miR-16-5p (Cur/miR-16-NBs)&#xff0c;a dual-functional platform rationally constructed to achieve synergism between PD-L1 suppression and immunogenic cell death (ICD) induction. The NBs were rationally engineered to exhibit spherical morphology. Only average particle size was quantified in this work, while polydispersity index and zeta potential data were not recorded, with quantitative characterization confirming high encapsulation efficiency of both miR-16-5p and curcumin. MiR-16-5p can downregulate the expression of PD-L1, inhibit the PD-1/PD-L1 signaling pathway. Meanwhile, as a safe and effective sonosensitizer, curcumin can generate abundant ROS under ultrasound irradiation to trigger ICD further promoting tumor cells to release three key signals-calreticulin (CRT), adenosine triphosphate (ATP), and high-mobility group box 1 protein (HMGB1), activate dendritic cells and boost CTL-mediated anti-tumor activity, thereby partially alleviating local tumor immunosuppression. Combined miR-16-5p and curcumin can activate intratumoral T lymphocytes to a certain degree and exert synergistic tumor-killing effects.Collectively, these findings highlight the potential of Cur/miR-16-NBs as an effective therapeutic platform for HCC, offering a precision medicine approach to improve treatment efficacy.

Haemophagocytic lymphohistiocytosis (HLH) is a life-threatening hyperinflammatory syndrome that may be triggered by infections, including brucellosis. Brucellosis-associated HLH is uncommon in children and may mimic malignancy or severe viral illness.We report a school-aged boy who presented with prolonged fever, pancytopenia, hepatosplenomegaly, hyperferritinaemia, hypertriglyceridaemia and hypofibrinogenaemia, fulfilling six HLH-2004 diagnostic criteria. Blood cultures confirmed Brucella species infection.Targeted antimicrobial therapy with gentamicin, doxycycline and rifampicin resulted in complete clinical and haematological recovery without corticosteroids, intravenous immunoglobulin or cytotoxic therapy.This case highlights the importance of recognising infection-triggered HLH in endemic regions and demonstrates that early treatment of the underlying infection may reverse the hyperinflammatory state.

This review summarizes recent findings on WOX transcription factors, highlighting their roles in development, stress response, and metabolic signaling in various species, along with their potential applications in agriculture. The WOX (WUSCHEL-related homeobox) family is known to coordinate various plant biological processes, including embryogenesis, apical and lateral meristem maintenance, regeneration, and vegetative and reproductive organ formation. Over the past five years, significant advances have expanded our understanding of WOX proteins, revealing novel interactors, downstream targets, and both transcriptional and post-transcriptional regulatory mechanisms. Notably, metabolic enzymes have been shown to modulate WOX gene activity, and the interplay between WOX genes and CLE (CLAVATA3 (CLV3)/EMBRYO SURROUNDING REGION-RELATED) peptides has been further clarified. Additionally, previously unknown functions have been discovered for many WOX family members across all phylogenetic clades, including roles in stress response. This review synthesizes recent findings on Arabidopsis WOX genes and their orthologs across diverse species, highlighting newly identified mechanisms that fine-tune WOX activity during plant development and environmental adaptation.

Biological activated carbon (BAC) filters are extensively employed in drinking water treatment plants (DWTPs) as a post-ozonation step to enhance water quality. However, the efficacy of BAC in removing ozonation transformation products (OTPs) remains inadequately characterized. This study comprehensively evaluated BAC performance by investigating dissolved organic matter (DOM) transformation and disinfection by-product formation potential (DBP FP) in samples collected from a full-scale DWTP utilizing the O3-BAC process. Results showed limited overall DOM removal by O3-BAC: 7.7 % for dissolved organic carbon and 15.0 % for fluorescent fractions. Molecular transformation analysis demonstrated that BAC filtration selectively targeted ozonation-derived saturated and oxidized molecules (characterized by H/Cw = 1.14 and O/Cw = 0.36) while allowing more unsaturated and aromatic OTPs to persist, confirming BAC's crucial role in determining the fate of OTPs and subsequent DBP FP. The BAC filtration achieved significant removal: 25.2 % for trihalomethanes (THMs), 47.4 % for haloaldehydes, 4.6 % for haloacetonitriles (HANs) and 47.0 % for halonitromethanes relative to O3 effluent levels. Spearman analysis revealed the characteristics of molecular formulas strongly associated with DBP FP. Specifically, haloketones and HAN FP-correlated formulas shared more reduced and less oxygenated structural characteristics compared to those associated with other DBP classes. The molecular mechanism underlying O3-BAC efficacy was elucidated: OTPs serve as key DBP precursors; BAC preferentially removes OTP-derived precursors (e.g., 89.1 % for THMs versus 5.8 % for original precursors); this selective removal counteracts ozonation's precursor generation effect. This synergy provides the molecular basis for DBP-FP reduction, clarifying BAC's critical role in controlling ozonation-derived risks.

Soil microplastics (MPs) pollution poses significant risks to environmental and human health, particularly in agricultural systems. However, limited data exist on MPs in farmland soils near mining regions, especially those associated with non-ferrous metal sulfide deposits. This study investigated the characteristics of MPs in agricultural soils surrounding sulfide mining areas. MPs were extracted by density separation, identified using &#xb5;FTIR, and statistically analyzed. Results revealed an average MP abundance of 19,610 n/kg, with dominant MPs (> 80 %) falling within the 0-100 &#x3bc;m size range. The most prevalent MP features included polystyrene, fragments, and transparent. Spatial analysis indicated a significant inverse correlation (P < 0.001) between MP abundance and the sample site distance to the mining area. We found that sulfide minerals decreased soil pH and oxidation-reduction potential (ORP), and that MPs abundance was positively correlated with ORP and negatively correlated with the concentrations of Fe, Cu, As. Agricultural activities, plastic waste disposal, and industrial activities jointly contributed to MP pollution. These findings highlight the need for integrated environmental management strategies to address co-occurring MPs and heavy metal pollution in mining-affected farmlands. Comprehensive monitoring of these pollutants is essential for developing targeted mitigation measures.

There has been an increased interest in the development and use of digital health technologies (DHTs) to help treat patients with opioid use disorder (OUD), but information on the needs and preferences of the patients seeking treatment for it is lacking. This study's objective was to collect data on preferences for DHTs that are intended to support patients with OUD. A discrete choice experiment was conducted with 500 respondents who self-reported a diagnosis of OUD. The experiment included 6 attributes identified based on earlier phases of the study. These attributes included the benefit of the DHT, data sharing with one's care team, data sharing with third parties, personalized goal tracking, community support, and health care support. Tools that allow patients to connect with their care team, connect with peers, or set and track their own goals were highly valued by respondents. In general, respondents who participated in the study preferred DHTs that improve the chance of abstaining from nonprescribed opioid use over those that improve the chance of staying in treatment or reducing the chance of opioid overdose. However, we also observed heterogeneity in preferences for benefits and data-sharing features within OUD treatment, suggesting that different patients may exhibit different preferences for OUD treatment in clinical settings. DHTs can be designed to target different outcomes and can include various features that support patients being treated for OUD. Although some outcomes and features emerged as being preferred by respondents, there was significant heterogeneity in the results. It will be important for developers to think about the various needs and preferences of target users (eg, patients) for their specific DHT rather than viewing the patient population with OUD as homogenous.

This article outlines a practical approach for general practitioners to evaluate and diagnose orthopedic conditions in small animals. Thorough history-taking for cases of lameness is reviewed, and the determination of onset, duration, severity, progression, and response to treatment is emphasized. Gait evaluation is described, which can identify the affected limb, lameness severity, and suspected neurologic involvement. Standing orthopedic examination techniques help assess symmetry and joint effusion. The recumbent exam is used to determine the source of the lameness through the assessment of pain, instability, and the application of targeted examination techniques and maneuvers such as the cranial drawer test.

Due to the rapidly evolving landscape of targeted therapies, there is an unmet need for comprehensive molecular profiling to guide treatment decisions for patients with large B-cell lymphoma (LBCL). Therefore, we designed a pilot study to assess the feasibility and turnaround time (TAT) of a comprehensive whole exome sequencing (WES) and transcriptome sequencing (RNA-seq) assay in patients with LBCL (NCT05464823). Patients aged &#x2265;18 years with LBCL were eligible. Formalin-fixed paraffin-embedded tissues at diagnosis or recurrence underwent WES, RNA-seq, and copy number analysis with concomitant germline DNA sequencing. Genomic and transcriptomic data were profiled to define various LBCL signatures such as cell-of-origin (COO), dark zone signature (DZsig), LymphGen and lymphoma microenvironment (LME) classification. Among 100 patients enrolled, samples from 71 patients (48 with newly diagnosed and 23 with relapsed/refractory disease) passed pathology quality control and underwent WES and RNA-seq analysis and reporting. The median TAT was 8 days for individual patient reports (range: 6-22 days), with 73% of reports delivered &#x2264;10 days. Applying molecular risk classification, high risk event-free survival within 24 months (EFS24) signature was associated with DZsig and LME, but not with COO or LymphGen indicating the complex heterogeneity of the disease. We demonstrated the clinical utility and acceptable TAT of a comprehensive WES and RNA-seq assay for LBCL managed in routine clinical practice. These findings support the real-world feasibility of using integrated WES and RNA-seq to define molecular subgroups, guide clinical decision-making at the time of a new line of therapy, and enable biology based subtype-driven clinical trials.