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bioRxiv [Preprint]. 2025 Jul 29:2025.07.24.666388. doi: 10.1101/2025.07.24.666388.

ABSTRACT

Protein phosphatase 2A (PP2A), an important therapeutic target, comprises a scaffold subunit PR65 composed of 15 HEAT (Huntingtin/elongation/A-subunit/TOR1) repeats, a catalytic subunit, and one of many different regulatory subunits that enable binding to specific substrates. Recently, small molecule activators of PP2A (SMAPs) were identified, although their mechanisms of action have not been fully defined. Here we explore the interaction of PR65 with two SMAPs, ATUX-8385 and the non-functional DBK-776, using single-molecule optical tweezers, ensemble methods, and computational analysis. In the absence of SMAP, PR65 shows multiple unfolding and refolding transitions, and the force-extension profiles are very heterogeneous with evidence of misfolding. Similar heterogeneity has been observed for chemical-induced unfolding of tandem-repeat proteins like PR65, a consequence of the internal symmetry of the repeat architecture. In the presence of ATUX-8385, higher unfolding and refolding forces are observed globally, and there is less misfolding, suggesting that ATUX-8385 acts like a pharmacological chaperone. In contrast, DBK-766-binding induces higher unfolding forces for only a few repeats of PR65, suggestive of a more localised effect; moreover, subsequent stretch-relax cycles show that PR65 is irreversibly locked in the unfolded state. Docking and molecular dynamics simulations provide additional insights how SMAP binding modulates PR65 structure and function.

PMID:40766424 | PMC:PMC12324213 | DOI:10.1101/2025.07.24.666388

bioRxiv [Preprint]. 2025 Jul 29:2025.07.24.666388. doi: 10.1101/2025.07.24.666388.

ABSTRACT

Protein phosphatase 2A (PP2A), an important therapeutic target, comprises a scaffold subunit PR65 composed of 15 HEAT (Huntingtin/elongation/A-subunit/TOR1) repeats, a catalytic subunit, and one of many different regulatory subunits that enable binding to specific substrates. Recently, small molecule activators of PP2A (SMAPs) were identified, although their mechanisms of action have not been fully defined. Here we explore the interaction of PR65 with two SMAPs, ATUX-8385 and the non-functional DBK-776, using single-molecule optical tweezers, ensemble methods, and computational analysis. In the absence of SMAP, PR65 shows multiple unfolding and refolding transitions, and the force-extension profiles are very heterogeneous with evidence of misfolding. Similar heterogeneity has been observed for chemical-induced unfolding of tandem-repeat proteins like PR65, a consequence of the internal symmetry of the repeat architecture. In the presence of ATUX-8385, higher unfolding and refolding forces are observed globally, and there is less misfolding, suggesting that ATUX-8385 acts like a pharmacological chaperone. In contrast, DBK-766-binding induces higher unfolding forces for only a few repeats of PR65, suggestive of a more localised effect; moreover, subsequent stretch-relax cycles show that PR65 is irreversibly locked in the unfolded state. Docking and molecular dynamics simulations provide additional insights how SMAP binding modulates PR65 structure and function.

PMID:40766424 | PMC:PMC12324213 | DOI:10.1101/2025.07.24.666388

Endoscopy. 2025 Aug 4. doi: 10.1055/a-2675-4322. Online ahead of print.

ABSTRACT

Background & Aims Post-endoscopic retrograde cholangiopancreatography (ERCP) pancreatitis (PEP) is common. Though multiple pancreatic duct (PD) cannulations are a known risk factor for PEP, the impact of single cannulations remains controversial. We aimed to identify whether single PD cannulation is associated with PEP. Methods We examined a prospective multi-center cohort of patients undergoing ERCP for biliary indications between 2021-2024, with third-party intra-procedural data recording and 30-day follow-up. PEP was defined using consensus definitions. Associations between PD cannulations and PEP were evaluated with multivariable logistic regression, with other patient- and procedure-related risk factors and preventive interventions used as covariates. Results were reported as odds ratios (ORs). Results PEP occurred in 282 (3.8%) of 7,430 ERCPs across nine centers. From multivariable analysis, PD cannulation was statistically significantly associated with PEP, with similar odds for single and multiple cannulations in first-time patients (OR=2.03, 95%CI [1.32-3.14] for single, OR=2.18 95%CI [1.18-4.00] for 5 or more cannulations). This was also true for all-comers (OR=1.97, 95%CI [1.33-2.93] for single, OR=2.15, 95%CI [1.21-3.82] for 5 or more). PD cannulation to the head (OR=2.09, 95%CI [1.36-3.21]) and body (OR=2.42, 95%CI [1.56-3.79]) were both associated with PEP, while side branch cannulations alone were not (OR=1.18, 95%CI [0.64-2.06]). Conclusions Single main PD duct canulation is independently associated with PEP and appears to be responsible for most of the magnitude of the association with PD cannulation. These data lend additional support to the use of preventative interventions such as PD stenting in cases where the PD is inadvertently cannulated.

PMID:40759180 | DOI:10.1055/a-2675-4322

Small. 2025 Aug 3:e06023. doi: 10.1002/smll.202506023. Online ahead of print.

ABSTRACT

Transition metal dichalcogenides are promising alternatives to noble metal catalysts, e.g., for (photo-)activation of greenhouse gases or hydrogenations. Herein, a direct synthetic route for 2D TiS2 nanosheets on Au(111) by titanium deposition in the presence of a mild, organic, non-oxidizing sulfur source is presented. High-resolution scanning tunneling microscopy (STM) is used to gain atomic-level insights into the TiS2 nanosheet morphology. In contrast to the literature, this protocol gains mostly hexagonal and truncated triangular nanosheets with an increased edge contrast in STM, analog to metallic edge states in MoS2. Synchrotron-based photoelectron spectroscopy allows insights into compositional details, specifically to distinguish different S sites on the TiS2 sheets and other S species on the sample. Further, a minimum size is identified (9 S atoms side length), which underlines the importance of moiré reconstructions for stress relief. The TiS2 sheets coexist with [Au]Ti1S3 clusters, in which a single gold atom is alloyed into the surface and capped by three S atoms. Together with the finding of a critical sheet size, this points toward on-surface Ostwald ripening as a relevant process in the sheet formation. Ab-initio calculations (density functional theory) underscore that the chemical potential of S is an essential descriptor to maintain shape control.

PMID:40754752 | DOI:10.1002/smll.202506023

Faraday Discuss. 2025 Aug 1. doi: 10.1039/d5fd90019k. Online ahead of print.

NO ABSTRACT

PMID:40748046 | DOI:10.1039/d5fd90019k

Children (Basel). 2025 Jun 20;12(7):812. doi: 10.3390/children12070812.

ABSTRACT

BACKGROUND/OBJECTIVES: Early care and education programs promote children's social-emotional development, predicting later school success. The COVID-19 pandemic worsened an existing youth mental health crisis and increased teacher stress. Therefore, we applied an infant and early childhood mental health consultation model, Jump Start Plus COVID Support (JS+CS), aiming to decrease behavioral problems in children post-pandemic.

METHODS: A cluster randomized controlled trial compared JS+CS to an active control, Healthy Caregivers-Healthy Children (HC2), at 30 ECE centers in low-income areas in South Florida. Participants were not blinded to group assignment. Teachers reported on children's social-emotional development at baseline and post-intervention using the Devereux Early Childhood Assessment and Strengths and Difficulties Questionnaire. We assessed whether teacher stress, classroom practices, and self-efficacy mediated the relationship between JS+CS and child outcomes. We also explored whether baseline behavior problems moderated JS+CS effects on child protective factors, relative to HC2.

RESULTS: Direct group-by-time differences between JS+CS and HC2 were limited. However, JS+CS demonstrated significant within-group improvements in teacher-reported child protective factors, behavior support practices, and classroom safety practices. Classroom safety practices consistently mediated positive changes in child behaviors, including the DECA total protective factor score and subdomains of initiative and self-regulation. Additionally, teacher perceptions of behavior support mediated gains in child attachment.

CONCLUSIONS: JS+CS shows promise in building protective systems around children through intentional support for teachers, underscoring the value of whole-child, whole-environment approaches in early intervention.

PMID:40723005 | DOI:10.3390/children12070812

J Gastroenterol Hepatol. 2025 Jul 28. doi: 10.1111/jgh.70031. Online ahead of print.

NO ABSTRACT

PMID:40717675 | DOI:10.1111/jgh.70031

Front Microbiol. 2025 Jul 10;16:1612534. doi: 10.3389/fmicb.2025.1612534. eCollection 2025.

ABSTRACT

The development and spread of antibiotic resistance in wastewater pose significant threats to both the environment and public health. Bacteria harboring multiple antibiotic resistance genes (ARGs), including those associated with horizontal gene transfer (HGT), can serve as persistent reservoirs and vectors for antimicrobial resistance in natural ecosystems. In this study, nine antibiotic-resistant bacterial strains (U1-U9) were isolated from a wastewater treatment plant (WWTP) effluent. The isolates were identified using 16S rRNA gene sequencing and whole-genome sequencing (WGS), and their antibiotic susceptibility profiles were evaluated. All isolates exhibited resistance to multiple antibiotics, and WGS revealed that U1, U2, U4, and U7 harbored diverse ARGs, including β-lactamase genes, efflux pumps, and resistance determinants for sulfonamides, tetracyclines, and, quinolones, confirming the presence of multidrug-resistant bacteria in WWTP effluent. Phylogenetic analysis classified them into Microbacterium spp. (Actinobacteria), Chryseobacterium spp. (Bacteroidetes), Lactococcus lactis spp. (Firmicutes), and Psychrobacter spp. (Proteobacteria). To explore mitigation strategies, eleven natural compounds were screened for their effects on cell growth, biofilm formation, and motility in selected multi-drug-resistant bacteria. Among the tested compounds, curcumin and emodin showed the most consistent inhibitory activity, particularly against Microbacterium spp. strains U1 and U2, and Lactococcus lactis sp. U4. In contrast, Chryseobacterium sp. U7, a Gram-negative strain, exhibited strong resistance to all tested natural compounds, highlighting the challenge of controlling Gram-negative ARBs in wastewater settings. These findings underscore the environmental risks posed by multidrug-resistant and HGT-associated ARG-harboring bacteria in WWTP effluent. They also demonstrate the potential of natural products, such as curcumin and emodin, as alternative or complementary agents for mitigating antibiotic resistance in water systems.

PMID:40708915 | PMC:PMC12286950 | DOI:10.3389/fmicb.2025.1612534

J Comp Physiol A Neuroethol Sens Neural Behav Physiol. 2025 Jul 24. doi: 10.1007/s00359-025-01752-7. Online ahead of print.

ABSTRACT

The naked mole-rat (NMR, Heterocephalus glaber) is a subterranean rodent that exhibits a range of unusual physiological traits, including diminished inflammatory pain. For example, nerve growth factor (NGF), a key inflammatory mediator, fails to induce sensitization of sensory neurons and thermal hyperalgesia in NMRs. This lack of NGF-induced neuronal sensitization and thermal hyperalgesia results from hypofunctional signaling of the NGF receptor, tropomyosin receptor kinase A (TrkA). Like NGF-TrkA signaling, the neurotrophic factor artemin, a member of the glial cell line-derived neurotrophic factor (GDNF) family, is implicated in mediating inflammatory pain through its receptor, GDNF family receptor α3 (GFRα3), which is expressed by a subset of dorsal root ganglia (DRG) sensory neurons. Here we investigated GFRα3 expression in DRG neurons of mice and NMRs, as well as measuring the impact of artemin on DRG sensory neuron function in both species in vitro. Using immunohistochemistry, we observed a similar abundance of GFRα3 in mouse and NMR DRG sensory neurons, high coexpression with the transient receptor potential vanilloid 1 (TRPV1) ion channel suggesting that these neurons are nociceptive neurons. Using in vitro electrophysiology to record from cultured DRG sensory neurons, we observed that artemin induced depolarization of the resting membrane potential and decreased the rheobase in both species, as well as diminishing the degree of TRPV1 desensitization to multiple capsaicin stimuli. Overall, results indicate that artemin similarly sensitizes sensory neurons in both mice and NMRs, future in vivo studies being required to confirm if the conserved in vitro sensitization also occurs in vivo.

PMID:40705105 | DOI:10.1007/s00359-025-01752-7

J Comp Physiol A Neuroethol Sens Neural Behav Physiol. 2025 Jul 24. doi: 10.1007/s00359-025-01752-7. Online ahead of print.

ABSTRACT

The naked mole-rat (NMR, Heterocephalus glaber) is a subterranean rodent that exhibits a range of unusual physiological traits, including diminished inflammatory pain. For example, nerve growth factor (NGF), a key inflammatory mediator, fails to induce sensitization of sensory neurons and thermal hyperalgesia in NMRs. This lack of NGF-induced neuronal sensitization and thermal hyperalgesia results from hypofunctional signaling of the NGF receptor, tropomyosin receptor kinase A (TrkA). Like NGF-TrkA signaling, the neurotrophic factor artemin, a member of the glial cell line-derived neurotrophic factor (GDNF) family, is implicated in mediating inflammatory pain through its receptor, GDNF family receptor α3 (GFRα3), which is expressed by a subset of dorsal root ganglia (DRG) sensory neurons. Here we investigated GFRα3 expression in DRG neurons of mice and NMRs, as well as measuring the impact of artemin on DRG sensory neuron function in both species in vitro. Using immunohistochemistry, we observed a similar abundance of GFRα3 in mouse and NMR DRG sensory neurons, high coexpression with the transient receptor potential vanilloid 1 (TRPV1) ion channel suggesting that these neurons are nociceptive neurons. Using in vitro electrophysiology to record from cultured DRG sensory neurons, we observed that artemin induced depolarization of the resting membrane potential and decreased the rheobase in both species, as well as diminishing the degree of TRPV1 desensitization to multiple capsaicin stimuli. Overall, results indicate that artemin similarly sensitizes sensory neurons in both mice and NMRs, future in vivo studies being required to confirm if the conserved in vitro sensitization also occurs in vivo.

PMID:40705105 | DOI:10.1007/s00359-025-01752-7

J Clin Tuberc Other Mycobact Dis. 2025 Jul 14;40:100552. doi: 10.1016/j.jctube.2025.100552. eCollection 2025 Aug.

ABSTRACT

BACKGROUND: Performance of OMNIgene.SPUTUM (OM-S) for transporting sputum was evaluated.

METHODS: This exploratory study was conducted during January-December 2019 at four near and one distant healthcare-facilities of Dhaka. Smear-positive pulmonary TB patients' sputa were collected, divided into 'OM-S untreated' and 'OM-S treated' portions, and transported to testing laboratory, Dhaka, on same-day from near-sites, and through courier from distant-site for smear-microscopy, culture, and Xpert MTB/RIF (Xpert) testing. Subset of 'OM-S treated' sample was tested with Xpert without centrifugation. Test results of all portions were compared in between.

RESULTS: Total 444 participants were enrolled (near-sites:198, distant-site: 246). All test results were comparable in both portions for near-sites. For distant-site, smear-microscopy's positivity was reduced by 4.1 % in 'OM-S treated', Xpert showed 100 % concordance in both portions, and culture was higher in 'OM-S treated' than 'OM-S untreated' (92.3 % vs 89.4 %; p = 0.288). Primary contamination rate in 'OM-S treated' was lower than 'OM-S untreated' (2.0 % vs 9.8 %; p < 0.05). For all sites, median (IQR) time-to-culture positivity was 35 (28, 42) days in both portions. Xpert positivity was 99 % concordant in 'OM-S treated' regardless of centrifugation.

CONCLUSIONS: OM-S is safe for sputum transportation. OM-S mixed sputum can be tested with Xpert and culture. Further studies can validate findings and assess cost-effectiveness.

PMID:40697678 | PMC:PMC12281178 | DOI:10.1016/j.jctube.2025.100552

Osteoarthritis Cartilage. 2025 Jul 19:S1063-4584(25)01091-X. doi: 10.1016/j.joca.2025.07.011. Online ahead of print.

ABSTRACT

OBJECTIVE: Osteoarthritis (OA) has a multifactorial pathogenesis, pain being the main symptom driving clinical decision making. Dysregulation of multiple mediators occurs in OA, the roles of many remaining to be identified. In dogs and humans with OA, synovial fluid lipidome dysregulation occurs, some findings being replicated in the plasma lipidome in a mouse OA model. One upregulated lipid is the sphingomyelin N-palmitoyl-D-erythro-sphingosylphosphorylcholine (d18:1/16:0), referred to here as SM. This study aimed to determine if SM causes joint pain and neuronal hyperexcitability in mice.

DESIGN: The effects of SM or a structurally related ceramide (CM) on mouse sensory neuron excitability were measured using patch-clamp electrophysiology, as well as the ability of intraarticular SM and CM to induce inflammatory pain in mice.

RESULTS: Incubation of sensory neurons with 1 µM SM decreased rheobase, compared to incubation with vehicle (p-adj = 0.0000146, 95% confidence interval (CI): 50.20, 76.73 pA) or CM (p-adj = 0.138, CI: 103.45, 171.55 pA). Similarly, SM induced mechanical hypersensitivity in mice compared to mice receiving vehicle (p-adj = 0.000003, 95% confidence interval (CI): 166.82, 251.63 g) or CM (p-adj = 0.055, 95% CI: 218.28, 268.12 g), which was coupled with a significant decrease in rheobase of knee-innervating neurons isolated from SM-injected mice compared to those receiving vehicle (p-adj = 0.0138, CI: 50.19, 76.73 pA) or CM (p-adj = 1.0, CI: 103.45, 171.55 pA).

CONCLUSIONS: The results generated demonstrate that a dysregulated lipidome can contribute to inflammatory OA pain, further work being necessary to determine the mechanism by which SM exerts its activity.

PMID:40691970 | DOI:10.1016/j.joca.2025.07.011

Osteoarthritis Cartilage. 2025 Jul 19:S1063-4584(25)01091-X. doi: 10.1016/j.joca.2025.07.011. Online ahead of print.

ABSTRACT

OBJECTIVE: Osteoarthritis (OA) has a multifactorial pathogenesis, pain being the main symptom driving clinical decision making. Dysregulation of multiple mediators occurs in OA, the roles of many remaining to be identified. In dogs and humans with OA, synovial fluid lipidome dysregulation occurs, some findings being replicated in the plasma lipidome in a mouse OA model. One upregulated lipid is the sphingomyelin N-palmitoyl-D-erythro-sphingosylphosphorylcholine (d18:1/16:0), referred to here as SM. This study aimed to determine if SM causes joint pain and neuronal hyperexcitability in mice.

DESIGN: The effects of SM or a structurally related ceramide (CM) on mouse sensory neuron excitability were measured using patch-clamp electrophysiology, as well as the ability of intraarticular SM and CM to induce inflammatory pain in mice.

RESULTS: Incubation of sensory neurons with 1 µM SM decreased rheobase, compared to incubation with vehicle (p-adj = 0.0000146, 95% confidence interval (CI): 50.20, 76.73 pA) or CM (p-adj = 0.138, CI: 103.45, 171.55 pA). Similarly, SM induced mechanical hypersensitivity in mice compared to mice receiving vehicle (p-adj = 0.000003, 95% confidence interval (CI): 166.82, 251.63 g) or CM (p-adj = 0.055, 95% CI: 218.28, 268.12 g), which was coupled with a significant decrease in rheobase of knee-innervating neurons isolated from SM-injected mice compared to those receiving vehicle (p-adj = 0.0138, CI: 50.19, 76.73 pA) or CM (p-adj = 1.0, CI: 103.45, 171.55 pA).

CONCLUSIONS: The results generated demonstrate that a dysregulated lipidome can contribute to inflammatory OA pain, further work being necessary to determine the mechanism by which SM exerts its activity.

PMID:40691970 | DOI:10.1016/j.joca.2025.07.011

Am J Cardiovasc Dis. 2025 Jun 15;15(3):175-180. doi: 10.62347/GLVD7571. eCollection 2025.

ABSTRACT

A coronary artery calcium (CAC) score of 0 is generally indicative of a low risk for both all-cause mortality and cardiovascular events, often serving as a basis for excluding obstructive coronary artery disease (CAD). Although isolated cases of coronary involvement have been reported in patients with a CAC score of 0, the incidence of extensive multivessel disease under these circumstances is exceedingly rare. A 48-year-old man with diabetes and hypercholesterolemia presented with atypical non-exertional left-sided chest pain. Despite a nonspecific ECG, a HEART score of 3, and a zero CAC score on echocardiography, coronary computed tomography angiography (CCTA) revealed multiple non-calcified plaques in the right coronary artery (RCA), right posterior descending coronary artery (RPDA), and left circumflex artery (LCX). The patient underwent staged percutaneous coronary intervention with drug-eluting stents, resulting in complete resolution of the stenosis. At the one-month follow-up, he remained symptom-free and tolerated the medication regimen well. This case report demonstrates that a zero CAC score should not preclude further evaluation in high-risk symptomatic patients. Extensive non-calcified plaques causing significant luminal obstruction underscore the limitations of CAC scoring, highlighting the need for additional imaging modalities, such as CCTA, to achieve timely and accurate diagnoses and appropriate therapeutic interventions.

PMID:40689064 | PMC:PMC12267082 | DOI:10.62347/GLVD7571

Drug Alcohol Depend. 2025 Jul 8;274:112774. doi: 10.1016/j.drugalcdep.2025.112774. Online ahead of print.

ABSTRACT

INTRODUCTION: Machine learning may enable detection of opioid misuse to prevent opioid-related risks including overdose and opioid use disorder.

METHODS: Here, we collected 9238 datapoints from on-body sensors and cognitive tasks in a sample of 169 patients who were prescribed opioid analgesics to manage chronic pain. We categorized patients into one of two groups using the Current Opioid Misuse Measure (COMM): those showing signs of opioid misuse (MISUSE+, n = 116) and those without signs of opioid misuse (MISUSE-, n = 53). Heart rate variability and respiration rate were assessed while participants completed a Dot Probe task involving shifting attention towards and away from opioid-related and emotional cues, and a Go/No-Go task involving inhibition of automatic responses. Cross-sectional data (e.g., physiological responses, task reaction times, task accuracy) were analyzed with a temporal fusion transformer machine learning (ML) model to predict COMM opioid misuse status. We employed Leave-One-Group-Out (LOGO) cross-validation with the participants divided into 10 groups. Each cycle, one group was held out for testing, ensuring robust, unbiased model validation across different subsets of participants.

RESULTS: The ML model showed good predictive performance for identifying opioid misuse (AUC, 0.81; specificity, 0.78; sensitivity, 0.78). Behavioral responses were stronger predictors of misuse status than physiological signals.

CONCLUSIONS: ML models using data from cognitive tasks and on-body sensors detected opioid misuse with an accuracy comparable to gold-standard self-reported opioid misuse assessments. Wearable sensors may provide only incremental predictive power over behavioral responses. Our ML model should be benchmarked against objective measures of opioid misuse.

PMID:40684523 | DOI:10.1016/j.drugalcdep.2025.112774

Br J Cancer. 2025 Jul 17. doi: 10.1038/s41416-025-03117-y. Online ahead of print.

ABSTRACT

BACKGROUND: BOADICEA is a widely used algorithm for predicting breast and ovarian cancer risks, using a combination of genetic and lifestyle, hormonal and reproductive risk factors. However, it has largely been developed using data from White/European individuals, limiting its applicability to other ethnicities. Here, we updated BOADICEA to provide ethnicity-specific risk estimates.

METHODS: We utilised data from multiple sources to derive estimates for the distributions and effect sizes of risk factors in major UK ethnic groups (White, Black, South Asian, East Asian, and Mixed), along with ethnicity-specific population cancer incidences. We also developed a method for deriving adjusted polygenic scores for individuals of mixed genetic ancestry.

RESULTS: The predicted average absolute risks were smaller in all non-White ethnic groups than in Whites, and the risk distributions were narrower. The proportion of women classified as at moderate or high risk of breast or ovarian cancer, according to national guidelines, was considerably smaller in non-Whites.

DISCUSSION: The updated BOADICEA, available in the CanRisk tool ( www.canrisk.org ), is based on more appropriate estimates for non-White women in the UK. Further validation of the model in prospective studies is required. Considering these findings, risk classification guidelines for non-White women may need to be revised.

PMID:40676226 | DOI:10.1038/s41416-025-03117-y

Spine (Phila Pa 1976). 2025 Jul 16. doi: 10.1097/BRS.0000000000005453. Online ahead of print.

ABSTRACT

STUDY DESIGN: retrospective analysis of prospectively collected data.

OBJECTIVE: to investigate whether two clustering approaches applied to the same database would lead to differences in the minimal clinical important difference (MCID) for health-related quality of life parameters (HRQoL).

SUMMARY OF BACKGROUND DATA: Machine learning approaches are being increasingly employed for the analysis of complex and heterogeneous settings such as that of adult spine deformity (ASD). However, it is not yet clear whether and how the choice of number and type of variables impacts the outcomes of a study.

METHODS: Two previously published clustering approaches (C12 and C16) were applied to a multicentric database of ASD patients who underwent surgery and had a minimum follow-up of one year. After clustering, the MCID for the Oswestry Disability Index, SRS-22, and SF-36 PCS were calculated for all clusters using the ROC method.

RESULTS: Data from 516 patients were available. Both algorithms led to a division of the database in three clusters, which presented similar characteristics both for C12 and C16. In particular, patients in clusters 1 to 3 presented an increasing level of imbalance and disability. The MCID for ODI, SRS-22, and SF-36 for each cluster differed between C12 and C16, but a similar pattern of increase of the MCID from Cluster 1 to Cluster 3 was observed for all HRQoL parameters and in both C12 and C16. The error rate, however, was smaller for C16.

CONCLUSION: Different clustering algorithms applied to the same database allowed to obtain similar clusters of ASD patients. However, the obtained MCIDs for the evaluated HRQoL parameters were different, highlighting the relevance of the choice of variables for the investigation of these parameters. The results suggest that clinically-driven clusters should be used when investigating clinical outcomes, as they allow for a smaller error rate.

PMID:40667701 | DOI:10.1097/BRS.0000000000005453

AoB Plants. 2025 Jun 26;17(4):plaf036. doi: 10.1093/aobpla/plaf036. eCollection 2025 Aug.

ABSTRACT

Legume introduction is effective for boosting primary productivity in C4-grass-dominated subtropical and tropical grasslands by overcoming nitrogen (N) limitation and consequently improving radiation-use efficiency (RUE), a key metric underlying plant production. However, an excessive proportion of C3 legumes may negatively affect RUE, especially in warm climates. We assessed the relationship between aboveground RUE and legume proportion of Paspalum notatum Flügge (C4 grass) mixtures with a tropical legume (Arachis glabrata Benth.; 0%-80%) under different defoliation and N fertilizer treatments in two studies over 3 years in Florida, USA. Linking the field data to a conceptual model, RUE was optimized at 26%-30% legume proportion across studies and years. When pastures were N-fertilized, RUE plateaued at 26% legume (0.60 g MJ-1) and linearly decreased with higher legume proportions. When pastures were unfertilized, RUE showed a quadratic relationship with legume proportion, being maximized at 30% legume (1.10 g MJ-1), overyielding the RUE in only-grass and legume-dominated sites by 110% and 86%, respectively. These responses suggest that RUE is N-limited when legume is below 30% in unfertilized canopies and is physiologically limited when legume is above 30% due to replacement of the C4 grass with a C3 legume. These findings provide a robust rationale to target low-to-moderate legume proportions in tropical grasslands for optimizing production and other ecosystem services. We empirically demonstrated that optimum legume proportion is ∼30% in a C4-grass-based tropical grassland compared with previous observations of ≥40% in C3-grass-based temperate grasslands, relevant insights for the design and maintenance of grassland ecosystems.

PMID:40667455 | PMC:PMC12260219 | DOI:10.1093/aobpla/plaf036

RSC Chem Biol. 2025 Jun 23. doi: 10.1039/d5cb00079c. Online ahead of print.

ABSTRACT

Accurate measurements of binding kinetics, encompassing equilibrium dissociation constant (K D), association rate (k on), and dissociation rate (k off), are critical for the development and optimisation of high-affinity binding proteins. However, such measurements require highly purified material and precise ligand immobilisation, limiting the number of binders that can be characterised within a reasonable timescale and budget. Here, we present the SpyBLI method, a rapid and cost-effective biolayer interferometry (BLI) pipeline that leverages the SpyCatcher003-SpyTag003 covalent association, eliminating the need for both binder purification and concentration determination. This approach allows for accurate binding-kinetic measurements to be performed directly from crude mammalian-cell supernatants or cell-free expression blends. We also introduce a linear gene fragment design that enables reliable expression in cell-free systems without any PCR or cloning steps, allowing binding kinetics data to be collected in under 24 hours from receiving inexpensive DNA fragments, with minimal hands-on time. We demonstrate the method's broad applicability using a range of nanobodies and single-chain antibody variable fragments (scFvs), with affinity values spanning six orders of magnitude. By minimising sample preparation and employing highly controlled, ordered sensor immobilisation, our workflow delivers reliable kinetic measurements from crude mixtures without sacrificing precision. We expect that the opportunity to carry out rapid and accurate binding measurements in good throughput should prove especially valuable for binder engineering, the screening of next-generation sequencing-derived libraries, and computational protein design, where large numbers of potential binders for the same target must be rapidly and accurately characterised to enable iterative refinement and candidate selection.

PMID:40655043 | PMC:PMC12247212 | DOI:10.1039/d5cb00079c

RSC Chem Biol. 2025 Jun 23. doi: 10.1039/d5cb00079c. Online ahead of print.

ABSTRACT

Accurate measurements of binding kinetics, encompassing equilibrium dissociation constant (K D), association rate (k on), and dissociation rate (k off), are critical for the development and optimisation of high-affinity binding proteins. However, such measurements require highly purified material and precise ligand immobilisation, limiting the number of binders that can be characterised within a reasonable timescale and budget. Here, we present the SpyBLI method, a rapid and cost-effective biolayer interferometry (BLI) pipeline that leverages the SpyCatcher003-SpyTag003 covalent association, eliminating the need for both binder purification and concentration determination. This approach allows for accurate binding-kinetic measurements to be performed directly from crude mammalian-cell supernatants or cell-free expression blends. We also introduce a linear gene fragment design that enables reliable expression in cell-free systems without any PCR or cloning steps, allowing binding kinetics data to be collected in under 24 hours from receiving inexpensive DNA fragments, with minimal hands-on time. We demonstrate the method's broad applicability using a range of nanobodies and single-chain antibody variable fragments (scFvs), with affinity values spanning six orders of magnitude. By minimising sample preparation and employing highly controlled, ordered sensor immobilisation, our workflow delivers reliable kinetic measurements from crude mixtures without sacrificing precision. We expect that the opportunity to carry out rapid and accurate binding measurements in good throughput should prove especially valuable for binder engineering, the screening of next-generation sequencing-derived libraries, and computational protein design, where large numbers of potential binders for the same target must be rapidly and accurately characterised to enable iterative refinement and candidate selection.

PMID:40655043 | PMC:PMC12247212 | DOI:10.1039/d5cb00079c