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The ROBINS-I tool is a commonly used tool to assess risk of bias in non-randomised studies of interventions (NRSI) included in systematic reviews. The reporting of ROBINS-I results is important for decision-makers using systematic reviews to understand the weaknesses of the evidence. In particular, systematic review authors should apply the tool according to the guidance provided. This study aims to describe how ROBINS-I guidance is currently applied by review authors. In January 2023, we undertook a citation search and screened titles and abstracts of records published in the previous 6 months. We included systematic reviews of non-randomised studies of intervention where ROBINS-I had been used for risk-of-bias assessment. Based on 10 criteria, we summarised the diverse ways in which reviews deviated from or reported the use of ROBINS-I. In total, 492 reviews met our inclusion criteria. Only one review met all the expectations of the ROBINS-I guidance. A small proportion of reviews deviated from the seven standard domains (3%), judgements (13%), or in other ways (1%). Of the 476 (97%) reviews that reported some ROBINS-I results, only 57 (12%) reviews reported ROBINS-I results at the outcome level compared with 203 reviews that reported ROBINS-I results at the study level alone. Most systematic reviews of NRSIs do not fully apply the ROBINS-I guidance. This raises concerns around the validity of the ROBINS-I results reported and the use of the evidence from these reviews in decision-making.

Cancer is the leading cause of death worldwide characterized by patient heterogeneity and complex tumor microenvironment. While the genomics-based testing has transformed modern medicine, the challenge of diverse clinical outcomes highlights unmet needs for precision oncology. As functional molecules regulating cellular processes, proteins hold great promise as biomarkers and drug targets. Mass spectrometry (MS)-based clinical proteomics has illuminated the molecular features of cancers and facilitated discovery of biomarkers or therapeutic targets, paving the way for innovative strategies that enhance the precision of personalized treatment. In this article, we introduced the tools and current achievements of MS-based proteomics, choice of discovery and targeted MS from discovery to validation phases, profiling sensitivity from bulk samples to single-cell level and tissue to liquid biopsy specimens, current regulatory landscape of MS-based protein laboratory-developed tests (LDTs). The challenges, success and future perspectives in translating research MS assay into clinical applications are also discussed. With well-designed validation studies to demonstrate clinical benefits and meet the regulatory requirements for both analytical and clinical performance, the future of MS-based assays is promising with numerous opportunities to improve cancer diagnosis, treatment, and monitoring.

Effective public health decision-making relies on rigorous evidence synthesis and transparent processes to facilitate its use. However, existing methods guidance has primarily been developed within clinical medicine and may not sufficiently address the complexities of public health, such as population-level considerations, multiple evidence streams, and time-sensitive decision-making. This work contributes to the European Centre for Disease Prevention and Control initiative on methods guidance development for evidence synthesis and evidence-based public health advice by systematically identifying and mapping guidance from health and health-related disciplines.Structured searches were conducted across multiple scientific databases and websites of key institutions, followed by screening and data coding. Of the 17,386 records identified, 247 documents were classified as 'guidance products' providing a set of principles or recommendations on the overall process of developing evidence synthesis and evidence-based advice. While many were classified as 'generic' in scope, a majority originated from clinical medicine and focused on systematic reviews of intervention effects. Only 41 documents explicitly addressed public health. Key gaps included approaches for rapid evidence synthesis and decision-making and methods for synthesising evidence from laboratory research, disease burden, and prevalence studies.The findings highlight a need for methodological development that aligns with the realities of public health practice, particularly in emergency contexts. This review provides a key repository for methodologists, researchers, and decision-makers in public health, as well as clinical medicine and health care in Europe and worldwide, supporting the evolution of more inclusive and adaptable approaches to public health evidence synthesis and decision-making.

The genus Medicago, which includes the widely cultivated forage crop alfalfa, is of significant agricultural and ecological importance. Understanding genetic diversity in Medicago is essential for the conservation of its germplasm and its utilisation in plant breeding. This study aimed to assess the genetic diversity and population structure of the Medicago germplasm collection at the German Federal Ex situ Gene Bank. Genotyping-by-sequencing was used to analyse 1234 accessions of the Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), representing 40 Medicago species. After filtering, a high-quality dataset of 23,315 single nucleotide polymorphisms (SNPs) was generated. Our analyses revealed distinct genetic clusters corresponding to Medicago species and sections, with cultivated M. sativa L. and M. × varia Martyn clustering together with less genetic diversity compared to their wild counterparts. This reflects the shared genetic composition and extensive gene flow between M. sativa and M. × varia, commonly considered a hybrid between M. sativa and M. falcata L. Wild species displayed a more complex genetic structure, with polyphyletic patterns indicating higher genetic differentiation that reflects their diverse evolutionary histories and ecological adaptations. In conclusion, the comprehensive diversity analysis of the IPK Medicago collection provides valuable insights for gene bank management, targeted conservation efforts and strategic breeding initiatives.

Calcium ions (Ca²⁺) are essential for plant development and stress responses, including heavy metal (HM) stress. However, the roles and mechanisms of calmodulin proteins (SlCalMs) in mediating cadmium (Cd) stress in Solanum lycopersicum, a model crop, remain poorly understood. This study aimed to investigate the calcium-mediated stress response in S. lycopersicum by identifying and characterizing the SlCalMs gene family, a key subfamily of calcium-binding proteins (CBPs), to elucidate their potential roles in stress tolerance. A genome-wide identification of SlCalMs was conducted using Oryza sativa sequences as a reference. Bioinformatics analyses included BLASTP searches, sequence alignment, phylogenetics, assessment of physicochemical properties, gene structure and motif analysis, chromosomal mapping and duplication events. Gene expression was assessed under Cd stress using RNA-seq and validated by quantitative real-time polymerase chain reaction (qRT-PCR). Molecular docking simulations evaluated Cd-binding affinities, and protein-protein interaction networks, and Gene Ontology (GO) enrichment were used to explore biological functions. Eight distinct SlCalM groups were identified, varying in gene size, exon number and isoelectric point. Conserved motifs, exon-intron patterns and stress-responsive cis-elements were identified. Chromosomal analysis revealed segmental duplications. Under Cd stress, several SlCalMs showed differential expression; notably, Solyc04g077830 was significantly downregulated and showed strong Cd-binding affinity in silico, suggesting a role in Cd sequestration. GO and interaction network analyses confirmed their involvement in Ca²⁺ signalling, metal ion binding and stress-related pathways. This study provides comprehensive insight into the structure, evolution and functional roles of SlCalMs in tomato. Their involvement in Ca²⁺ signalling and Cd stress response highlights their potential for improving HM tolerance, offering valuable targets for future genetic or biotechnological interventions in crop improvement.

Mass spectrometry imaging (MSI) technologies are widely used today to study the in situ spatial distributions for a variety of analytes. As these technologies advance, the pursuit of higher resolution in MSI has intensified. The limitation of direct desorption/ionization is its insufficient ionization, posing a constraint on the advancement of high-resolution MSI technologies. The introduction of postionization process compensates the low ionization efficiency caused by sacrificing the desorption area while pursuing high spatial resolution, resolving the conflict between high spatial resolution and high sensitivity in direct desorption/ionization method. Here, we discuss the sampling and ionization steps of MSI separately, and review the postionization methods in MSI according to three different sampling modes: laser sampling, probe sampling, and ion beam sampling. Postionization technology excels in enhancing ionization efficiency, boosting sensitivity, mitigating discrimination effect, simplifying sample preparation, and expanding the scope of applicability. These advantages position postionization technology as a promising tool for biomedical sciences, materials sciences, forensic analysis and other fields.

Mediterranean temporary ponds (MTPs) are dynamic habitats where low levels of dissolved oxygen can significantly impact plant life. This study investigated the effect of hypoxia and near-anoxia on seed germination and the induction of secondary dormancy in 14 plant species, characteristic of this habitat. Imbibed seeds were subjected to various oxygen concentrations (0.1, 5, 10, or 21% O₂), in both light and darkness. We also tested seed ability to recover germination by moving them to aerobic conditions. We measured embryo growth after hypoxic treatments and during recovery in three species with morpho-physiological dormancy, a rarely investigated response in this dormancy class. Our findings revealed a wide range of species-specific responses. Hypoxia did not inhibit germination in half of the tested species in the light, while near-anoxia completely inhibited germination in all species. However, most seeds fully recovered germination ability once aerobic conditions were restored. Interestingly, hypoxia in darkness reduced or prevented germination in some species and specifically induced secondary dormancy in Juncus bufonius. Surprisingly, seeds of Bulliarda vaillantii lost their light requirement for germination under hypoxia. In three Ranunculus species with morpho-physiological dormancy, hypoxia slowed embryo growth, which delayed germination recovery. This study reveals that MTPs species have evolved adaptations, ranging from tolerance to hypoxic conditions, to the ability to trigger secondary dormancy, which are crucial to surviving and reproducing in these unique environments. The results offer new insights into the germination ecophysiology of MTPs species and their regeneration niche in temporary wetlands.

Soybean (Glycine max), is an important oilseed crop that plays a vital role in ensuring global food security. However, it is susceptible to multiple abiotic stresses that can reduce yield. The ubiquitination-proteasome pathway is a crucial regulatory mechanism that controls a broad range of processes in plants. We investigated the function of Glycine max drought-induced SINA (GmDIS1), an E3 ligase gene, in soybean abiotic stress tolerance using Agrobacterium-mediated transformation to develop soybean GmDIS1-RNAi transgenic lines. GmDIS1 was significantly induced under drought and heat stress. Several physiological traits revealed resilience of GmDIS1-RNAi lines under drought and heat stress. The functions of stress-related genes, such as AOS and GmPAL were investigated to dissect the pathways that contribute to drought and heat tolerance in GmDIS1-RNAi lines. The results suggest that decreasing expression of GmDIS1 can enhance soybean tolerance to drought and heat, and also provide a significant target for developing more drought- and heat-tolerant soybean varieties and other crops.

Little is known about the functional role of silicon (Si) in metal-hyperaccumulating plant species, such as Arabidopsis halleri. We investigated the responses of A. halleri from two accessions, Bestwig (Best) and Langelsheim (Lan), to Si supplementation and insect infestation in two controlled full-factorial experiments. Plants were grown in soil either unsupplemented (-Si) or supplemented (+Si) with Si. Some of these plants were kept either uninfested or infested by larvae of the leaf beetle Phaedon cochleariae. Shoot chemical and mechanical traits and plant resistance against the larvae were quantified. Detached leaves from the remaining plants were used to examine whether trichome density and leaf area consumed by larvae were influenced by the accession and/or Si. We found that Si supplementation, but not insect infestation or their interaction, led to twice as high concentrations of shoot Si in +Si in comparison to -Si plants. Insect relative growth rate was not impacted by Si, but by accession, namely lower when larvae fed on Lan than on Best plants. Likewise, leaf area consumed by larvae was consistently lower in the former accession. The density of trichomes was twice as high in plants of the Lan than the Best accession. Uninfested +Si plants contained the highest C/N in both accessions. The composition of glucosinolates differed between accessions, with some glucosinolates being induced by Si, insect infestation or both in the Best plants only. Our findings highlight distinct (induced) defence strategies within A. halleri plants, which may indicate different local adaptations of the source populations.

Cyphomandra betacea, a valuable understory crop in southwestern China, exhibits high sensitivity to water availability. Under global climate change with increasingly erratic precipitation, understanding how Cyphomandra betacea, seedlings respond to rainfall variations is crucial for sustaining this distinctive industry. Through controlled experiments, this work systematically investigates how different rainfall patterns affect seedling growth and physiology, providing a theoretical basis for science-based management under future climate scenarios. Seedlings were subjected to a four-month simulated rainfall experiment with two rainfall intervals (T: 3-day; T₊: 6-day) and three rainfall amounts (W: control; W₊: +40%; W_-: -40%). Biomass, non-structural carbohydrates (NSC), and carbon, nitrogen, phosphorus stoichiometric characteristics were analysed. Seedling growth is more sensitive to variations in rainfall amount, and appropriate increases in rainfall can promote seedling growth and development. Under changes in rainfall patterns, seedlings prioritize the storage of NSC in stems, followed by leaves, with the lowest allocation to roots. Nitrogen content within organs is pivotal for the composition of NSC and can regulate the sugar-starch conversion process. The July W₊T treatment resulted in optimal performance for the majority of growth indicators and demonstrated the highest nutrient accumulation efficiency. We identified a stem-preferential carbon allocation strategy and systemic N limitation, offering key insights for conservation and cultivation under changing climates.