Advancing Open Science

Background: Platelets have conventionally been viewed as cellular fragments crucial for hemostasis; nonetheless, their extensive secretome of cytokines and growth factors has been increasingly acknowledged as a significant regulator of inflammation and tissue healing at the ocular surface. Aims: The objective of this narrative review is to synthesize existing knowledge of platelet biology with new findings about the therapeutic use of platelet-derived products in dry eye disease (DED). Methods: A qualitative review of the PubMed, Scopus, and Web of Science databases up to June 2025 identified preclinical, translational, and clinical studies assessing platelet-rich plasma (PRP), plasma rich in growth factors (PRGF), platelet lysate, and autologous serum tears for dry eye disease (DED) and associated ocular surface disorders. Results: Platelet-derived formulations have exhibited reliable immunomodulatory and regenerative effects by diminishing inflammatory signaling, lowering cytokine expression, and facilitating epithelial and neurotrophic restoration. Clinical investigations have indicated enhancements in tear film stability, corneal staining, and patient-reported symptoms, especially in cases of moderate-to-severe or refractory illness. Nonetheless, methodological diversity, inconsistent preparation techniques, and restricted sample sizes have impeded comparability among experiments. Conclusions: Platelet-derived treatments constitute a biologically viable and clinically promising strategy for the management of dry eye disease (DED). Future research must emphasize the standardization of preparation protocols, the identification of predictive biomarkers such as transforming growth factor-β1 (TGF-β1), nerve growth factor (NGF), and matrix metalloproteinase-9 (MMP-9), as well as the design of multicenter randomized controlled trials to guarantee reproducible, GMP-compliant clinical applications.

Background: Antimicrobial resistance is a growing global health threat, with beta-lactamases playing a central role in resistance to beta-lactam antibiotics. Building on our previous survey of 2340 putative beta-lactamases, we conducted an in-depth analysis of 129 prioritized candidates (70–98.5% amino acid identity to characterized enzymes) detected in 102 bacterial genera across 13 phylogenetic classes from environmental, animal, and human sources worldwide. Methods: We applied a motif-centric assessment of class-defining catalytic residues, evaluated the genomic context using a heuristic Index of Proximal Mobility (IPM) derived from the two immediately adjacent open reading frames, and examined the phylogenetic placement. AI-based substrate predictions were generated at a restricted scope as exploratory evidence. Results: Candidates spanned all Ambler classes (A–D); preservation of catalytic motifs was common and consistent with potential catalytic activity. Twelve of 129 (9.3%) loci had nearby mobile-element types (e.g., insertion sequences, integrases, transposases) and scored High IPM, indicating genomic contexts compatible with horizontal gene transfer. We also observed near-identical class A enzymes across multiple genera and continents, frequently adjacent to mobilization proteins. Conclusions: We propose a reproducible, bias-aware, early warning framework that prioritizes candidates based on motif integrity and mobility context. The framework complements existing surveillance (GLASS/EARS-Net) and aligns with a One Health approach integrating human, animal, and environmental reservoirs. Identity thresholds and IPM are used for inclusion and contextual prioritization, rather than as proof of function or mobility; AI-based predictions serve as hypothesis-generating tools. Experimental studies will be essential to confirm enzymatic activity, mobility, and clinical relevance.

Approximately two-thirds of athletes who are submitted to Anterior Cruciate Ligament Reconstruction (ACLR) never return to their preinjury level of performance, potentially due to muscle strength deficiencies or altered loading patterns during landing or jumping tasks. This study aimed to estimate individual muscle forces during a double-leg drop jump task, and assess sagittal plane between-limb asymmetries in muscle forces and ground reaction forces using a musculoskeletal modelling approach, in athletes who underwent ACLR. Thirty male field-sport athletes (age: 18–35 years; mass: 84.3 ± 12.3 kg; height: 180.2 ± 8.4 cm) post-ACLR (39.8 ± 3.9 weeks) using patellar or quadriceps tendon grafts were tested. Scaled musculoskeletal models were implemented in OpenSim, and muscle forces were estimated using the Computed Muscle Control optimization method. The contralateral limb exhibited greater vertical ground reaction forces across most of the rebound phase (d = 2.01). Compared with the contralateral limb, the ACLR limb showed reduced quadriceps (d = 1.72), soleus (d = 0.95), and gluteus maximus (d = 0.83) forces, indicating deficits in knee extensor, plantarflexor, and hip extensor neuromuscular function. Smaller asymmetries were found for the gluteus medius (d = 0.60) and hamstrings (d = 0.72), while other muscles showed symmetrical activation patterns. These results reveal persistent between-limb asymmetries in muscle recruitment and loading up to nine months post-ACLR, emphasizing the importance of targeted rehabilitation to restore symmetrical neuromuscular control during explosive movements.

Parkinson’s disease is a neurodegenerative disorder disease that worsens over time and involves the deterioration of nerve cells in the brain. Gait analysis has emerged as a promising tool for early detection and monitoring of Parkinson’s disease. However, the accurate classification of Parkinsonian gait is often compromised by missing body keypoints, particularly in critical regions like the hip and legs that are important for motion analysis. In this study, we propose RecovGait, a novel method that combines a gated initialization technique with unscented tracking to recover missing human body keypoints. The gated initialization provides initial estimates, which are subsequently refined through unscented tracking to enhance reconstruction accuracy. Our findings show that missing keypoints in the hips and legs significantly affect the classification result, with accuracy dropping from 0.8043 to 0.5217 in these areas. By using the gated initialization with an unscented tracking method to recover these occluded keypoints, we achieve an MAPE value as low as 0.4082. This study highlights the impact of hip and leg keypoints on Parkinson’s disease gait classification and presents a robust solution for mitigating the challenges posed by occlusions in real-world scenarios.

Platelet shortage poses a significant barrier to research and transfusion therapies because native megakaryocytes (MKs) are scarce in blood. To overcome this limitation, pluripotent stem cell–derived MKs (PSC-MKs) offer a standardized, donor-independent platform for research and therapeutic development, including disease modeling and ex vivo platelet production. Here, we report a chemically defined, feeder-free protocol to generate MKs from human pluripotent stem cells (hPSCs). The protocol combines the small molecule MPL agonist Butyzamide, macrophage colony-stimulating factor (M-CSF), and three-dimensional (3D) suspension culture, achieving high efficiency and reproducibility. Butyzamide replaced recombinant thrombopoietin (TPO), yielding comparable CD41⁺/CD42b⁺ populations and enhanced polyploidization. M-CSF accelerated nuclear lobulation and induced 4N MKs, while 3D culture increased yield, cell size, and substrate detachment. Multiple independent assays confirmed mature MK hallmarks, multi-nuclei, demarcation membranes, granules, and elevated mitochondrial respiration. Single-cell RNA sequencing outlined a continuous trajectory from early progenitors to functionally specialized MK subsets. This platform enables reliable MK supply for mechanistic studies and in vitro platelet production, advancing both basic research and therapeutic development.

Accurately assessing grazing impacts is essential for sustaining alpine grasslands. Conventional approaches often rely on total forage productivity, an indirect and uncertain proxy for forage availability. In this study, we propose a novel framework for estimating grazing pressure that integrates residual biomass with grazing intensity, thereby overcoming the limitations and uncertainties inherent in total forage-based assessments. Our results reveal pronounced spatiotemporal variation in grazing intensity: lowland areas experienced the highest intensity early in the growing season, whereas upland areas became more heavily grazed later in the season. However, grazing intensity alone proved insufficient to explain grazing pressure or predict pasture degradation risk. Overlay analyses demonstrated that only 38.8% of high intensity areas identified as under high grazing pressure, and more than 40% of high intensity area exhibiting substantial aboveground biomass. These findings highlight the limited explanatory power of grazing intensity when considered in isolation. By explicitly incorporating standing biomass rather than relying merely on total production, the proposed framework reduces estimation uncertainty, enhances ecological realism, and provides a scalable, more accurate and practical tool for monitoring grassland utilization and degradation.

The Yellow River Basin serves as a critical ecological barrier and economic corridor in China, playing a pivotal role in national ecological security and sustainable development. This study develops a comprehensive evaluation framework grounded in the Water–Energy–Food–Ecosystem (WEFE) nexus, employing 25 indicators across nine provinces and autonomous regions over the period 2000–2023. Utilizing a multi-method approach—including the entropy weight method, coupling coordination degree model, center of gravity migration analysis, principal component analysis, and obstacle factor diagnosis—the research investigates the coordinated development and dynamic interactions among the WEFE subsystems. Key findings include: (1) the calculated weights of the water, energy, food, and ecological subsystems were 0.3126, 0.1957, 0.1692, and 0.3225, respectively, indicating that ecological and water subsystems exert the greatest influence; (2) distinct growth patterns among subsystems, with the energy subsystem exhibiting the fastest growth rate (212%) and the water subsystem the slowest (4%); (3) a steady improvement in the overall coordination degree of the WEFE system, rising from 0.417 in 2000 to 0.583 in 2023—a 39.8% increase—with Henan (0.739) and Inner Mongolia (0.715) achieving the highest coordination levels in 2023, while Qinghai (0.434) and Ningxia (0.417) remained near imbalance thresholds; (4) complex spatial dynamics reflected by cumulative center of gravity migration distances of 678.2 km (water), 204.9 km (energy), 143.3 km (food), and 310.9 km (ecology) over the study period; and (5) identification of per capita water resources as the principal limiting factor to coordinated WEFE development, with an obstacle degree of 0.1205 in 2023, underscoring persistent water scarcity challenges. This integrated framework advances WEFE nexus analysis and provides robust, evidence-based insights to inform regional policy and resource management strategies.

The Middle East respiratory syndrome coronavirus (MERS-CoV) remains a highly significant threat to global public health. Dromedary camels are the zoonotic source of human infection. All cases of zoonotic Middle East respiratory syndrome (MERS) have occurred in Middle Eastern countries despite MERS-CoV infection of camels being widespread in Africa. This disparity in the geographic burden of the disease may be due to genomic differences between MERS-CoV circulating in Middle Eastern countries (clades A and B) versus those infecting camels in Africa (clade C), although the precise genetic determinants of virulence remain to be elucidated. The objective of the studies reported here was to evaluate differences in the magnitude of virus shedding and in transmissibility of clades A/B and C viruses using alpacas as a surrogate for dromedary camels. We found that two of three African-origin, clade C strains of MERS-CoV induced very reduced levels of virus shedding and were transmitted inefficiently to contact control animals as compared to one other clade C virus and representative viruses from clade A and B. Lower virus titers in the nasopharynx may be associated with lower zoonotic transmission and human disease severity and may explain the observed epidemiology of MERS-CoV in Africa where zoonotic disease appears rare. These results add to our understanding of the transmission of different lineages of MERS CoV in camelids and zoonotic transmission.