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The quantification of protein levels is an important part of standard cerebrospinal fluid (CSF) analysis. However, delays in processing due to shipment and/or laboratory opening schedules, combined with the need to preserve sample stability during storage, pose critical challenges to ensuring accurate diagnoses. Our aim was to assess the stability of CSF proteins in canine and feline samples refrigerated and subsequently frozen. CSF samples were prospectively collected from dogs and cats. CSF protein levels were measured: immediately after collection; after refrigeration (at 4°C for 48 h); and after freezing (at -20°C for 6 to 150 days). Linear regression was performed for each species to evaluate the influence of time and preservation on protein levels. The impact of the variation in CSF protein measurements on clinical interpretation was evaluated. Fourteen dogs and four cats with neurological diseases, and five cats euthanized for reasons unrelated to neurological issues were enrolled in the study. Protein measurements were stable over time in dogs (p = 0.334) and cats (p = 0.785), regardless of the preservation method and timing of analysis. The variation between the CSF protein measurements of fresh and stored samples did not change the interpretation of any results. CSF protein levels in dogs and cats remained stable when samples were refrigerated for 48 h prior to being frozen for long-term storage, supporting the feasibility of banking samples for research and laboratory control. Additional studies are necessary to evaluate the stability of specific protein biomarkers.

Companion animal brain banking has been recognized as a valuable approach for translational aging and dementia research. However, realizing the full value of canine brain banks depends on optimizing the methods that are used to collect and preserve the tissue. Whole brain perfusion fixation is one promising approach, but it is not yet well described in dogs. Here we describe the development of methods for a canine brain bank (currently n = 55), including whole brain perfusion fixation via aortic cannulation and brain extraction. We assessed perfusion quality using gross examination, post-perfusion CT, and histological clearance of blood vessels. We found that body weight and average flow rate per body weight were each significantly correlated with perfusion quality in our cohort. To illustrate the kind of analysis the bank could facilitate, we next performed a preliminary study of brain aging, one of our primary planned research applications. Using a pixel classifier applied to whole slide images, we quantified lipofuscin burden, and in this preliminary cohort found that it increased strongly with age in both the thalamus and hippocampus. In the hippocampus, lipofuscin burden was also elevated in dogs with owner-reported cognitive dysfunction, although the current cohort is too small to determine to what extent this association is independent of age. Preliminary electron microscopy studies also confirmed that perfusion fixed tissue from the bank is amenable to ultrastructural analysis. This work describes one approach for canine brain perfusion fixation and introduces a brain tissue resource that may help support future neuroscience research.

Somatic cells serve as essential genetic resources for biobanking. Traditionally, jaguar fibroblast-like cells are frozen in cryovials using slow-freezing protocols. However, this approach necessitates access to ultra-low temperature freezers and requires substantial storage capacity. This study aimed to evaluate the efficacy of a rapid-freezing protocol as a feasible alternative for the freezing of jaguar fibroblasts. Following cultivation to the third passage, cells were frozen using either slow-freezing (SF) or rapid-freezing (RF). For SF, cells were frozen in cryovials (-1 &#xba;C/min), whereas RF involved rapid-freezing (-60 &#xba;C/min) using nitrogen vapor and storage in 0.25 mL straws. Post-thaw, cells were re-cultured to assess recovery. Cell viability was assessed via light microscopy (Trypan blue), fluorescence microscopy (acridine orange + propidium iodide), and flow cytometry (Hoechst 33342 + propidium iodide) immediately post-thaw and upon reaching confluency (> 80%). Fresh culture viability assessed by Trypan blue, acridine orange, and flow cytometry was 98.2 &#xb1; 0.3%, 88.7 &#xb1; 3.9%, and 94.2 &#xb1; 2.2%, respectively. No significant differences in viability were observed between SF and RF immediately post-thaw or after subsequent cultivation. All three assays revealed significant differences (p < 0.05) between cells assessed immediately after thawing and those assessed at confluence. Upon reaching confluence, there was no significant difference (p > 0.05) between fresh and frozen-thawed cells. These findings indicate that RF in straws is a viable alternative to SF for jaguar fibroblast cryopreservation, offering optimized storage density for genetic resource banks.

The SUPREMO trial reported adjuvant chest wall radiotherapy had no effect on 10-year overall survival (primary endpoint) in patients with intermediate-risk breast cancer after mastectomy. The quality of life (QOL) substudy of SUPREMO (UK patients only) examines the effects of chest wall radiotherapy in patients with intermediate-risk breast cancer 1 year, 2 years, 5 years, and 10 years after treatment. Here, we report 5-year QOL results (a secondary endpoint), including prespecified subgroup analyses. SUPREMO is an international, parallel-group, randomised, controlled, phase 3 trial. Women aged 18 years or older with intermediate-risk breast cancer (defined as pT1-2N1 or pT3N0 or pT2N0 with histological grade 3, or lymphovascular invasion, or both) who had undergone mastectomy (and axillary surgery) were randomly assigned (1:1) to receive chest wall radiotherapy (40-50 Gy in 15-25 fractions) or no radiotherapy. Randomisation was in permuted blocks with varying block length, stratified by centre, without masking of patients or investigators. All UK patients were invited to the QOL substudy. Consenting patients completed validated questionnaires (European Organisation for Research and Treatment of Cancer Quality of Life Core 30-item questionnaire; European Organisation for Research and Treatment of Cancer Quality of Life Questionnaire-Breast Cancer Module, 23 items; Body Image Scale; and Hospital Anxiety and Depression Scale) before randomisation and at 1 year, 2 years, 5 years, and 10 years. Here, we report the prespecified primary QOL outcomes of global QOL, fatigue, physical function, chest wall symptoms, shoulder and arm symptoms, body image, anxiety, and depression in the intention to treat population, using repeated measures mixed-effects models. This trial is registered with the ISRCTN registry, ISRCTN61145589 and is now completed. Between Aug 4, 2006, and April 29, 2013, SUPREMO recruited 1691 patients internationally, of whom 1679 were randomly assigned to receive chest wall radiotherapy (n=845) or not (n=834). Of 1233 patients from UK centres (619 in the chest wall radiotherapy group and 614 in the no chest wall radiotherapy group), 989 (80%) consented to the QOL substudy: 947 (96%) of 989 returned baseline questionnaires and 620 (75%) of 832 patients returned the expected year-5 questionnaires. Chest wall symptoms were worse with chest wall radiotherapy (effect estimate 1&#xb7;99 [95% CI 0&#xb7;36-3&#xb7;62]; p=0&#xb7;017) with an improvement between years 1, 2, and 5. Chemotherapy was associated with less improvement over time (effect estimate 2&#xb7;97 [95% CI 0&#xb7;24-5&#xb7;71]; p=0&#xb7;033) but without interaction with chest wall radiotherapy. No statistically significant differences were observed in other QOL domains (arm symptoms, body image, fatigue, pain, overall QOL, physical functioning, or anxiety and depression). In the subgroup analyses, patients who had a sentinel lymph node biopsy followed up by an axillary lymph node clearance with chest wall radiotherapy had worse chest wall symptoms than those having a single axillary operation (difference chest wall radiotherapy vs no chest wall radiotherapy -5&#xb7;29 [95%CI -8&#xb7;53 to -2&#xb7;05]; p=0&#xb7;0015). The 5-year QOL findings showed worse chest wall toxicity and no effect on other QOL domains. The subgroup analyses suggested a potential negative effect of chemotherapy and axillary clearance after sentinel node biopsy. UK Medical Research Council, National Institute for Health and Care Research, European Organisation for Research and Treatment of Cancer, Edinburgh Trustees of Breast Cancer Institute in Edinburgh and the Edinburgh Cancer Centre NHS Endowment funds, The Dutch Cancer Society, Trustees of the Hong Kong and Shanghai Banking Corporation.

Successful pregnancies with healthy newborns have been reported after all types of solid organ transplantation (SOT). Limited data in young children suggest similar development and health compared with the general population. However, fetal development may be influenced by factors such as immunosuppressive medication and the increased incidence of pregnancy complications, and forthcoming health problems may only become apparent later in the life of the offspring. To allow for better investigation of the long-term health of offspring born after SOT, we designed the Transplantlines Next Generation biobank and cohort study. This study will be the first with detailed data on overall health status at later age in offspring born to mothers after kidney, liver, pancreas (including pancreas islet), heart and lung transplantation (KTx, LiTx, PTx, HTx and LuTx, respectively). Transplantlines Next Generation is a descriptive, prospective cohort study and biobank. It includes offspring aged&#x2265;16 years born after KTx or LiTx. Because of the scarcity of pregnancies after (solo) PTx, HTx and LuTx worldwide, and as a consequence the lack of information about these pregnancies, we want to include all offspring, at any age, born after (solo) PTx, HTx and/or LuTx in the Netherlands. Participants will attend a one-time visit for questionnaires, physical tests (for participants&#x2265;16 years including kidney ultrasound and 24-hour ambulatory blood pressure) and biological sample collection (urine, blood, faeces) for participants&#x2265;16 years of age for biobanking. Reference values and values from existing birth cohorts will serve as controls. Primary endpoints are cardiovascular and kidney health, assessed through growth charts, physical tests (eg, body composition, blood pressure, kidney ultrasound) and metabolic and kidney function biomarkers. Secondary aims include immunological status, microbiome analysis, quality of life and overall development. Ethical approval has been obtained from the local medical ethical committee; the Institutional Review Board METc UMC Groningen (METc 2023/610). The study will be conducted according to the Declaration of Helsinki and in accordance with the Medical Research Involving Human Subjects Act and other guidelines, regulations and Acts including the General Data Protection Regulation (GDPR). All participants will give written informed consent upon enrolment. TransplantLines Next Generation is designed to deliver pioneering insights into the health of offspring born after SOT. This knowledge will help optimise care and available information for families with a pregnancy wish after SOT and provide a rationale for future studies. The results of this study are planned to be submitted for publication in relevant peer-reviewed journals and will be presented at national and international conferences. NCT07291258.

Cyanobacteria-mediated synthesis of silver nanoparticles (AgNPs) offers a sustainable approach for producing functional AgNPs for biomedical applications. However, controlling their physicochemical properties while maintaining robust biological activities remains a critical challenge. Here, we developed a green, one-pot synthesis method to produce uniform, small AgNPs using the novel Cyanobium gracile PCC 6307 and evaluated their biological activities. PCC 6307 was isolated and identified using microscopic examination and sequencing analysis. The aqueous extract of algal biomass (CB) and cell-free medium (CS) were used to synthesize AgNPs under optimized reaction conditions. Acetyleugenol (38.09%) and eugenol (10.5%) were identified as the predominant metabolites in CB extract by gas chromatography-mass spectrometry. CS@AgNPs and CB@AgNPs showed strong absorption peaks at 412.5 and 430.5&#xa0;nm and exhibited uniform spherical shapes with core diameters of 4.0 and 12.4&#xa0;nm, respectively. The synthesized AgNPs exhibited hydrodynamic diameters&#x2009;<&#x2009;100&#xa0;nm and negative surface charges. Energy-dispersive X-ray spectroscopy confirmed the elemental composition of AgNPs, while Fourier-transform infrared spectroscopy verified the presence of algal functional groups on their surfaces. AgNPs reduced the colon cancer cell viability to 4.1% at 200&#xa0;&#xb5;g/mL exhibiting moderate to low toxicity toward normal human fibroblasts. CS@AgNPs revealed higher biocidal activity (inhibition zone&#x2009;&#x2264;&#x2009;20&#xa0;mm) than CB@AgNPs against Staphylococcus aureus, S. haemolyticus, and S. lentus. The antioxidant activity of AgNPs was assessed using the 2,2-diphenyl-1-picrylhydrazyl assay, where AgNPs showed moderate (46.0%) to low (25.1%) radical scavenging activity. This study provides a sustainable approach for producing uniform, small, and biologically active AgNPs with potential biomedical applications.

Cardiovascular-kidney-metabolic (CKM) syndrome emphasizes the interplay between metabolic dysfunction, kidney injury, and cardiovascular abnormalities. The triglyceride-glucose (TyG) and its obesity-related derivatives have emerged as simple biomarkers for insulin resistance, yet their association with aortic stenosis (AS) risk in CKM syndrome stages 0-3 remains unclear. This prospective cohort study included 363,907 participants with CKM stages 0-3 from the UK Biobank. We calculated six TyG-related indices (TyG, TyG-BMI, TyG-Waist, TyG-WHR, TyG-WHtR, and TyG-ABSI) and evaluated their associations with incident AS using Cox proportional hazards and competing risk models. Mediation analysis assessed the role of systolic blood pressure (SBP), and incremental predictive value was evaluated using C-index and net reclassification index (NRI). During 15.25 years of follow-up, 3,326 AS cases occurred. After full adjustment, each 1-SD increment in TyG-related indices was associated with 8-24% increased AS risk. Compared with the lowest tertile, the highest tertile showed 1.13-1.77-fold increased risk. All indices demonstrated linear dose-response relationships with AS. SBP mediated 12.1-18.1% of these associations. TyG-WHtR showed the greatest predictive improvement (C-index: 0.806; NRI: 0.308). TyG-related indices, particularly TyG-WHtR, are independently associated with AS risk in CKM syndrome stages 0-3, offering potential for early risk stratification and targeted prevention strategies.

Early detection of colorectal polyps is crucial to reduce the morbidity and mortality associated with colorectal cancer. However, during endoscopy, continuous camera motion and the complex clinical environment often degrade key visual cues (e.g., polyp morphology, texture, and boundaries). This leads to cross-frame view shifts, which are characterized by heterogeneous appearances of the same lesion over time, thereby introducing spurious correlations that hinder reliable assessment. To address this, we propose a causality-inspired representation learning framework with spatiotemporal memory for polyp detection in colonoscopy videos (CIRL-Polyp). Specifically, we develop a novel View-Shift-Aware Causal Intervention Module (VACIM) to remove non-causal influences by enforcing prediction invariance under view shift perturbations. To further constrain non-causal factors, we introduce a dual-branch detection framework that processes the original and intervention frame sequences in parallel and enforces prediction consistency across branches, thereby promoting invariance to non-causal variations. In addition, we propose Causal Temporal Consistency Memory (CTCM) to leverage long sequence-dependency features and stabilize causal representations by constructing memory banks across branches and performing temporal consistency-enhanced cross-attention. Comprehensive experiments on two public video datasets and a private dataset demonstrate that CIRL-Polyp outperforms existing methods, which validates the effectiveness and suggests the clinical potential of the proposed framework from a causal perspective.

Progressive supranuclear palsy (PSP) is a tauopathy for which there is limited understanding of epidemiological risk factors. The objective of this study was to identify premorbid nonmedical and medical risk factors for PSP diagnosis in a large prospective population-based cohort. We performed a matched nested case-control study using the UK Biobank (UKB) cohort consisting of >500,000 adults aged 37 to 73&#x2009;years. PSP diagnoses were identified from 2007 to censor date (August 2025) and matched 1:10 by age and sex to control subjects without parkinsonism or dementia diagnoses. We derived risk factors using an iterative logistic regression approach. There were 240 incident PSP diagnoses in the UKB. In the fully adjusted model, premorbid depression (odds ratio [OR], 3.22; 95% confidence interval [CI], 2.30-4.50; P&#x2009;<&#x2009;0.001), delirium (OR, 6.76; 95% CI, 4.08-11.19; P&#x2009;<&#x2009;0.001), and functional gastrointestinal disorders (OR, 1.91; 95% CI, 1.39-2.63; P&#x2009;<&#x2009;0.001) were associated with increased risk. Heavy alcohol consumption was associated with higher risk compared with moderate consumption (OR, 1.65; 95% CI: 1.21-2.26; P&#x2009;=&#x2009;0.0017). Body mass index (BMI) and alcohol consumption had nonlinear risk profiles. Cancer diagnosis was inversely associated with PSP (OR, 0.56; 95% CI, 0.40-0.79; P&#x2009;<&#x2009;0.001). Depression and delirium diagnoses remained significant 5 to 10&#x2009;years before diagnosis. In this large prospective cohort, PSP was associated with premorbid lifestyle practices and neuropsychiatric diagnoses years before diagnosis. This suggests a long premotor phase and highlights opportunities for earlier diagnosis and further mechanistic investigation. The inverse association with cancer mirrors the relationship seen in other neurodegenerative conditions and may point toward shared mechanisms. &#xa9; 2026 International Parkinson and Movement Disorder Society.

Vitamin K is essential for processes that inhibit vascular and valvular calcification, attributes of atherosclerotic cardiovascular diseases (ASCVDs) and aortic stenosis (AS). Relationships between dietary vitamin K1 and K2 intakes, ASCVD and AS were explored. In 112,111 participants from the UK Biobank without ASCVD or AS (median age 59.7 years, 57.3% female), dietary vitamin K1 (&#x3bc;g/day) and K2 (&#x3bc;gPKeq/day) intakes were calculated from Oxford WebQ 24-h dietary questionnaires, using vitamin K food composition databases. Associations with incident ASCVD, ASCVD subtypes and AS, obtained from self-reports, primary care data, hospital records and death registries, were examined using restricted cubic splines within multivariable-adjusted Cox proportional hazards models. Over a median follow-up of 10.5 years, 8696 ASCVD cases were recorded. Non-linear associations between vitamin K1 intakes and ASCVD were observed within multivariable-adjusted models (pnon-linearity&#x2264;0.01). Higher vitamin K1 intakes were associated with a 7-10% lower risk of ASCVD, with no advantage observed beyond moderate intakes (HRQ3vsQ1 0.90 [0.85-0.95]). Lower risk relationships were also found for ischemic heart disease (IHD), ischemic stroke and peripheral arterial disease but not AS. The highest vitamin K2 intakes were associated with a greater risk of incident ASCVD (HRQ5vsQ1 1.07 [1.01-1.14]), IHD and AS. Among adults without prevalent ASCVD, those with higher dietary vitamin K1, but not K2, intakes had lower ASCVD risk. Positive associations with vitamin K2 may reflect food source confounding warranting further investigation. These findings reinforce the promotion of vitamin K1-rich vegetable consumption (e.g. spinach and broccoli) to support cardiovascular health.

Slickwater fracturing fluid has gained wide attention in hydraulic fracturing due to its advantages, such as low pipeline friction and minimal formation damage. However, its low viscosity leads to poor sand-carrying performance, affecting the effectiveness of fracturing operations. There is an urgent need to develop a new slickwater system with an excellent sand-carrying performance. In this study, cellulose nanofibrils (CNFs) were modified via alkylation to prepare silane-modified nanofibers (denoted as S-CNFs). The successful preparation of S-CNFs was confirmed by Fourier transform infrared (FT-IR) spectroscopy and X-ray photoelectron spectroscopy (XPS). The research results indicate that S-CNFs have an excellent dispersion stability. S-CNFs were then introduced into the conventional slickwater to develop the S-CNFs-reinforced slickwater, which meets industry standards in terms of drag reduction and core damage evaluation. Through rheological tests, the S-CNFs-reinforced slickwater shows excellent strength and temperature shear resistance. Additionally, the system also shows the wider elasticity-dominated range in the viscoelasticity test, confirming the improvement in proppant-carrying capacity. Compared with the conventional slickwater, the S-CNFs-reinforced slickwater transports proppants deeper into fractures in the dynamic proppant-carrying experiment. It reduces the area of sand banks in simulated fractures by 16.4% and the residual sand amount by 14.8%. Scanning electron microscopy (SEM) was employed to observe the formation of the double-network structure constructed by polymers and S-CNFs in the S-CNFs-reinforced slickwater. This double-network structure confirms the intrinsic mechanism of the enhanced sand-carrying performance. These findings exhibit that S-CNFs can significantly improve rheological properties and sand-carrying capacity, providing a theoretical foundation for the further development of modified nanofiber-based slickwater fracturing fluids.

Migrant and left-behind children experience health vulnerability that are shaped by climate change, including by climate change, including illness, displacement, and family separation, but also by the organisation of healthcare systems. This scoping review maps the evidence on healthcare services for migrant and left-behind children, with attention to health needs, service provision, barriers and facilitators to access, and gaps in care. Following PRISMA-ScR reporting guidance, searches were conducted in Web of Science/MEDLINE and Scopus for peer-reviewed English-language publications from the database inception to May 2026. After duplicate removal and two-stage screening, 35 publications were included. The evidence shows that migrant and left-behind children require a continuum of preventive, primary, specialist, developmental, nutritional, dental, mental health, and emergency care. However, access is frequently constrained by legal exclusion, documentation requirements, cost, language barriers, limited health system navigation, discrimination, mobility, disrupted continuity of care, and under-resourced services. Left-behind children remain especially under-represented, with limited evidence on preventive, mental health, and chronic care access. Facilitators include inclusive entitlements, community outreach, school provision, interpreters, bilingual professionals, culturally responsive care, health literacy support, and cross-sector coordination. The review argues that healthcare inequity for children affected by migration is produced not only by service absence, but by systems that fail to adapt to children's legal, linguistic, relational, and transnational realities. Rights-based child health policy should therefore move from episodic and discretionary provision towards universal entitlements, continuous care pathways, and integrated support that is culturally responsive, developmentally appropriate, and accountable to the lived experiences of migrant and left-behind children.

ObjectiveThere is limited prospective evidence on the association of physical activity with metabolic dysfunction-associated steatotic liver disease. We aimed to use data from two major databases to systematically quantify the correlation of physical activity with the risk of metabolic dysfunction-associated steatotic liver disease.MethodsThis study used data from the National Health and Nutrition Examination Survey (2007-2020) and UK Biobank. Initially, a cross-sectional study was conducted using the data from the National Health and Nutrition Examination Survey data to assess the links between physical activity and metabolic dysfunction-associated steatotic liver disease using weighted logistic regression analysis with appropriate sample weights to ensure national representativeness. Subsequently, Cox proportional hazards regression and restricted cubic splines models were applied to infer causality and evaluate the potential dose-response relationships in the UK Biobank cohort over a median follow-up period of 14.18 years. Additionally, subgroup and sensitivity analyses were conducted to test the robustness of the results.ResultsIn the National Health and Nutrition Examination Survey, higher physical activity levels were inversely associated with metabolic dysfunction-associated steatotic liver disease risk (odds ratio: 0.78, 95% confidence interval: 0.63-0.97). This finding was supported in the UK Biobank cohort (hazard ratio: 0.57, 95% confidence interval: 0.34-0.76). A nonlinear dose-response relationship existed between physical activity and metabolic dysfunction-associated steatotic liver disease (p&#x2009;=&#x2009;0.023 for a nonlinearity). Subgroup analyses highlighted stronger effects in overweight/obese individuals (hazard ratio: 0.62, 95% confidence interval: 0.43-0.88), suggesting that this high-risk population may derive particular benefit from increased physical activity.ConclusionsAnalyses of two large databases demonstrated a nonlinear dose-response association between higher physical activity levels and reduced metabolic dysfunction-associated steatotic liver disease risk. Moderate physical activity levels (600 to <3000 metabolic equivalent of task-min/week) were significantly protective, and high physical activity levels (&#x2265;3000 metabolic equivalent of task-min/week) conferred additional benefits.

This article describes how the National Human Milk Bank (NHBM) of Israel handled challenges caused by the invasion of Southern Israel by Hamas on October 7, 2023, leading to the massacre of more than 1,200 Israeli civilians and the taking of more than 250 hostages into the Gaza Strip, including women and children. This created a need for donor human milk (DHM) for infants, not based on infants' medical conditions (routine usage), but rather because of maternal death, disability, or emergency deployment. We studied satisfaction levels among the families involved with the donated milk. We hypothesized that caretakers would experience initial anxiety related to providing infants DHM before its administration, but high levels of satisfaction following it. At the onset of the war, it was decided that the criteria for receiving DHM would be widened to include infants of mothers who were victims of the attack, infants injured in the attack, and infants of mothers emergently called up for military combat. Later on, criteria were expanded to include breastfeeding women psychologically impacted by the attack or its resultant displacement. Satisfaction was determined from questionnaires that were sent to families 6 months to 1 year after the events took place. Forty-six infants of 44 mothers (2 sets of twins) received DHM between October 2023 and January 2025. The most frequent reasons for eligibility were maternal deployment to active duty (n&#x2009;=&#x2009;20), maternal secondary exposure to events (n&#x2009;=&#x2009;18), and maternal death or severe injury (n&#x2009;=&#x2009;7). Twenty families returned the questionnaires, showing that at enrollment, approximately 50% of families expressed a high or moderate degree of anxiety. In contrast, after the infant received DHM, less than 25% still expressed anxiety. More than 80% attested that DHM donation helped them in handling the consequences of the war and contributed to their child's health, while more than 90% felt that the DHM donation contributed to their own well-being. In settings of maternal loss or incapacity during war, donor human milk can be effectively provided by human milk banks and lead to a high level of satisfaction. We suggest that human milk banks should be considered a component of emergency preparedness planning.