Lund University

Aging is accompanied by progressive physiological declines in body functions. However, the molecular mechanisms of aging remain poorly understood in decapod crustaceans, a diverse group of invertebrates. We investigated age-related differences in the hemolymph proteomes of marbled crayfish using label-free protein quantification (LC-MS/MS), applying a significance threshold of p<0.05 and a fold change >2 to elucidate molecular mechanisms underpinning aging. Two groups including young (n=6) and old (n=7) crayfish were used. Results showed a downregulation of superoxide dismutase in the hemolymph of older individuals. At the same time, glutathione peroxidase and transketolase were upregulated, which may reflect age-dependent changes in oxidative stress regulation and potential compensatory responses. Aging crayfish exhibited changes in levels of the ProPO system and phagocytosis-related proteins that suggest a possible shift from melanization to phagocytosis in the aging immune system. Additionally, the lower levels of some other immune-related proteins in the old individuals may be consistent with a decline in the crayfish immune system with age. Proteins associated with wound healing and regeneration were higher in young individuals, which may suggest an age-based decline in regenerative capacity. Cytoskeletal and extracellular matrix proteins were upregulated in older crayfish, which could potentially influence immune cell functions. Age-related alterations in the quantities of vitellogenins, hemocyanins, and metabolic enzymes maybe associated with changes in reproductive investment, respiratory capacity, and energy metabolism. Together, these findings highlight the complex molecular basis by which aging reshapes the hemolymph composition and alters immune system characteristics in an invertebrate, revealing molecular signatures that may represent aging mechanisms. Data are available via ProteomeXchange with identifier PXD065434.

Conventional methods of pea protein extraction are resource-intensive, involving multiple purification steps that remove co-existing components such as starch. In contrast, we investigate how minimally processed yellow pea protein extracts that retain residual carbohydrates form thermally induced gels. Contrast-variation small-angle neutron scattering, rheology, and differential scanning calorimetry show that salt-extracted samples form a unified gel matrix through co-gelation of protein and carbohydrate components, as evidenced by a shared contrast match point in the final network. Ultra-small-angle neutron scattering reveals mesoscale heterogeneity, with structural features persisting up to 2 μ m, indicative of a particulate, protein-rich network within the composite gel. Rheological and calorimetric analyses suggest that gelation proceeds via a kinetic pathway in which protein aggregation precedes unfolding, ultimately shaping network architecture and viscoelastic properties. These findings demonstrate that minimal processing can yield structurally integrated protein–starch gels and clarify how composition and processing determine network architecture and mechanical performance. • We study the thermal gelation of pea protein and carbohydrates from truncated extraction. • Co-gelation is evidenced via contrast-variation small-angle neutron scattering. • Model fitting and data-driven decomposition evidence multi-scale structures. • The gelation process is further monitored by rheology and calorimetry. • Protein-carbohydrate co-gelation seems promising for sustainable protein extraction.

There is limited data regarding the rare and aggressive colorectal neuroendocrine carcinoma (CR-NEC). In this large prospective study, molecular-clinical characteristics and treatment outcomes following palliative chemotherapy are reported for 163 metastatic CR-NEC patients, with a comparison to a population-based prospective cohort of 263 metastatic colorectal adenocarcinoma (CR-AC) patients. Eighty-three percent of CR-NEC received first-line platinum-etoposide, while 98% of CR-AC patients received first-line fluorouracil-based chemotherapy. Disease control rate across all first-line regimens in CR-NEC and CR-AC was 43% vs. 74%, immediate progressive disease 46% vs. 15%, progression-free survival 2.4 months (m) (95% CI 2.1-3.3) vs. 7.7 m (95% CI 6.9-8.5), and overall survival 6.7 m (95% CI 5.6-8.8) vs. 16.8 m (95% CI 13.7-20.3), all, p < .001. CR-NEC more often had synchronous metastases, worse performance status, and symptom burden at treatment initiation than CR-AC (all, p < .001). Two-year survival was 9% vs. 37% in CR-NEC and CR-AC (p < .001). BRAF mutations were frequent in CR-NEC and CR-AC (26% vs. 20%, p = .153) and associated with shorter OS in CR-NEC and CR-AC (p = .025 and p = .003). KRAS mutations were less frequent in CR-NEC than CR-AC (34% vs. 45%, p = .041), but only associated with shorter OS in rectal NEC (p = .04). The frequencies of APC and TP53 mutations were similar between the cohorts and did not impact survival. Metastatic CR-NEC and CR-AC are clinically distinct, with NEC demonstrating more aggressive features, limited treatment effect, and worse prognosis. Although they share important driver mutations, the underlying reason for their marked clinical differences remains unclear.

Timely discharge of patients who are clinically ready to be discharged from hospitals to the next point of care is a common health system challenge. Ensuring safe and effective discharge holds new urgency, given the backlogs and increased waiting times for inpatient services in many countries after the COVID-19 pandemic. The study aimed to identify policy options for addressing delayed discharges in six European countries (Denmark, France, Germany, the Netherlands, Norway, and Sweden), and to summarise available evidence on their effectiveness. Experts from the Health Systems and Policy Monitor (HSPM) network of the European Observatory on Health Systems and Policies and additional country experts identified relevant policies and initiatives up to November 2023. When evaluations were available, the experts also provided information on their findings. The data collection was followed by a qualitative, cross-country comparative analysis. A total of 17 policies or initiatives were identified in the study countries. Hospital-based initiatives included discharge planning and transitional care. Community and home care initiatives included municipal emergency beds, intensive home rehabilitation, and assisted discharge. Cross-sectoral initiatives ranged from coordination efforts at the regional and municipal levels to decision support systems and financial incentives. Several common factors or principles underpin many of the identified policies and initiatives. These include clarity of responsibility, effective planning and communication, resourcing of community-based capacity, possible unintended consequences of financial penalties, and the need to adopt a systemic approach.

Autistic persons experience problems with movements and body awareness in everyday functionality, expressed in affected movement quality. Yet, there is a knowledge gap about the effects of interventions targeting these aspects in autistic persons. To evaluate effects of Basic Body Awareness Therapy (BBAT) and treatment-as-usual (TAU), compared to TAU alone on movement quality in autistic persons. In this pragmatic RCT, autistic persons with no intellectual disorder, 15-30 years old and in contact with habilitation services, were randomized to an intervention group (IG) (n=28, 22 females) or a control group (CG) (n=29, 21 females). The IG received 12 sessions of individual BBAT and TAU, the CG received TAU. Primary outcome was a person-reported measure of an individualized movement quality problem, using a numeric rating scale (NRS). Secondary outcome was Body Awareness Scale Movement Quality (BAS MQ), i.e., scored physiotherapist observations. Between- and within-group analyses, and individual one-sided t-tests were performed. Comparing the between-group changes in NRS and BAS MQ after 12 weeks, the differences were statistically significant in favor of the IG (p<0.001). Within-group analyses: the IG had statistically significant improvements on both outcomes (p<0.001), the CG had not. Individually in the IG, 54% of the participants improved significantly in NRS and 41% in BAS MQ. In the CG, the corresponding numbers were 10% and 7%, respectively. BBAT and TAU were superior to TAU alone for improving movement quality. BBAT is a relevant intervention for autistic young adults to improve everyday functionality. Clinical Trials NCT0323876.

The growing use of crime scene dogs to detect human body fluids has led to an increased number of outdoor samples being submitted to forensic DNA laboratories. The analysis of DNA in outdoor samples such as soil presents significant challenges with present protocols due to the combination of relatively large sample volumes and co-extraction of humic substances. To address this, we have optimised a soil microbial DNA extraction protocol for the recovery of semen by incorporating enzymatic, chemical and physical lysis steps. This optimisation increased the DNA yield 43-fold. Using mock crime scene samples, the protocol provided high quality STR profiles from semen in 10mL of different types of soil (mixed forest soil, sandy soil and park soil), organic matrix (dry leaves) and inorganic matrices (sand and gravel). The protocol also generated complete STR profiles for soil samples containing blood, saliva and cell-free DNA. However, all humic substances were not removed, causing inhibition in MPS analysis. The extracted DNA consisted of more than 95% double-stranded (ds) DNA with equivalent fragment sizes as DNA extracted using magnetic bead-based kits. The protocol is thus compatible with library preparation methods that demand dsDNA, e.g. in whole genome sequencing. The protocol has been successfully implemented in casework and used in serious crime investigations. Human STR profiles have been generated from crime scene items in 50% of the cases. In the majority of these (82%), no results were generated using swabs and traditional DNA extraction methods.

• Stakeholders identified five themes for home-based stroke rehabilitation. • Patient involvement and staff competence were both rated feasible and important. • The physical home environment and social networks were rated as important, yet less feasible to implement. • A gap was found between policy ambitions and real-world rehabilitation delivery. • Concept mapping enabled stakeholder-driven priorities in care development. Despite growing emphasis on home-based rehabilitation following stroke, there remains limited understanding of how to effectively tailor services to individual needs. This study employed concept mapping to explore diverse stakeholder perspectives on adapting home-based rehabilitation to better support the recovery process after stroke. Through this mixed-methods participatory approach, we engaged 89 stakeholders, including stroke survivors, significant others, healthcare professionals, managers, and architects. Participants generated, sorted, and rated statements about what matters most in home-based stroke rehabilitation. A hierarchical cluster analysis was conducted to organize the data into concepts. Five clusters were identified: 1) social support at home and beyond; 2) professional competence; 3) supportive home environment; 4) teamwork and resources; and 5) patient involvement. Patient involvement was rated highest for importance and feasibility, followed by professional competence. Specifically, stakeholders emphasized that rehabilitation should feel meaningful and be tailored to the patient's needs. No generated ideas from the cluster “supportive home environment” were found in the go-zone, and adapting rehabilitation to diverse home environments was recognized as crucial but hard to implement. The findings underscore the need for individualized, context-sensitive approaches and suggest that strengthening professional competence, particularly in navigating diverse environments and engaging support networks, will be essential to translating person-centered principles into everyday rehabilitation practice.

Pea albumins are the soluble fraction of pea proteins, recovered as a side stream of the isoelectric precipitation. The time-resolved heat-induced structural transitions of the whole fraction of pea albumin (PA) were studied by evaluating the structure of the two main fractions, PA1 and PA2. Synchrotron radiation circular dichroism (SR-CD) spectroscopy showed that the secondary structure signature for PA1 remained unchanged at temperatures up to 85°C. In contrast, PA2 underwent a change in secondary structure between 45 and 70°C. These thermal transitions were also confirmed by Nano Differential Scanning Calorimetry (Nano-DSC). Small-angle X-ray scattering (SAXS) in situ experiments were also carried out, and high-resolution structural models of PA1 and PA2, derived from the AlphaFold Protein Structure Database, were used for data analysis. PA1 maintained a consistent local structure with a radius of gyration ∼17 Å across all temperatures, but formed small soluble aggregates above 60°C, as evidenced by an upturn at low q . PA2 scattering resulted from its dimeric structures and transitioned to unfolded structures after 60°C. The unfractionated PA scattering showed good agreement with a linear combination of PA1 and PA2 scattering. This was also confirmed by the structural behavior of the purified PA1+PA2 mixture, which closely resembled that of PA2, though the PA2 component remained partially folded even after heating above 70°C. This study brings new knowledge on the contribution of the two main components of pea albumins in the thermal behavior of this novel extract, with great potential to be used as a functional food ingredient. • PA1 and PA2 contains higher β-sheet compared to α-helix; • Heat induced aggregation of PA can be modeled after a combination of PA1 and PA2; • PA1 maintains a consistent local structure, but forms soluble aggregates above 60°C; • PA2 transitions from dimer to unfolded structure above 60°C.

The stability of therapeutic proteins is crucial for maintaining their efficacy and safety. Misfolded or aggregated proteins can accelerate aggregation by a seeding effect, altering protein stability and interactions. Heat-induced aggregates can pose a risk because they can have perturbed structures that expose aggregate-prone regions. This study explored seeding by heat-induced protein aggregates on the stability and aggregation of two therapeutic proteins, human growth hormone (hGH) and a monoclonal Antibody (Antibody A), during storage of two weeks at three relevant temperatures of 4 °C, room temperature (RT, 21-23 °C), and 40 °C. Grwoth of aggregates were seen in the sub-micron-sized aggregates by Dynamic Light Scattering at 4 °C or RT. Furthermore, heated Antibody A showed signs of reversibility of higher-order structure after storage. Samples stored at 4 °C and RT showed similar stability and aggregation behaviour. While two weeks of storage at 40 °C accelerated the structural degradation of both proteins, it did not accelerate aggregation or particle formation for either hGH or Antibody A.