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Using RT-QuIC seeding assays determine the presence of pathogenic alpha-synuclein (aSyn) aggregates in the submandibular gland (SMG), skin, and CSF from autopsy-confirmed cases of incidental Lewy body disease (ILBD), Parkinson disease (PD), and controls. Submandibular gland, skin, and CSF samples from autopsied cases in the Arizona Study of Aging and Neurodegenerative Disorders (AZSAND) were assayed using RT-QuIC methods. Correlations with the Unified Staging System for Lewy Body Disorders (USSLB) was performed. A total of 19 controls, 40 ILBD, and 15 PD cases were studied. While aSyn seeding assays were positive in all 3 tissues from ILBD cases, the sensitivity was much lower in the SMG (13/37, 35.1%) and skin (6/16, 37.5%) than in CSF (25/33, 75.8%). In PD sensitivity was good in all tissues: SMG (7/8, 87.5%), skin (7/8, 87.5%), and CSF (10/10, 100%), as was specificity in CSF (9/12, 75%), skin (8/9, 88.9%), and SMG (7/9, 77.8%). USSLB stage correlated with aSyn SAA positivity in all 3 tissues with CSF most sensitive to stages I and II (IIa and IIb) cases. In the 20 cases that had all 3 tissues tested only 2 were positive in the SMG but not CSF and one in skin but not CSF. There were 6 ILBD cases that had positive CSF but not SMG or skin, and one of these ILBD cases was USSLB stage I (olfactory bulb only). In this small study of autopsy-confirmed ILBD, aSyn seeding assays had moderately high sensitivity (75.8%) in the CSF but not in skin or SMG. In PD, the skin, SMG, and CSF showed high sensitivity, with specificity being similar in all tissues, although sample sizes were small. These results, although preliminary, suggest that detecting aSyn using seeding assays of CSF, but not the skin or SMG, may be valuable for identifying individuals with prodromal Lewy body disease. This is especially true for ILBD cases that often had Lewy bodies in the olfactory bulb only (USSLB Stage I) or in brainstem regions (Stage IIa), suggesting that detecting these cases may require CSF and not biopsies of the skin or SMG.

Background/Objectives: Spinal muscular atrophy (SMA) is a rare genetic disorder caused by mutations or deletions in SMN1, resulting in the loss of SMN protein and severe neuromuscular consequences. Nusinersen, an antisense oligonucleotide that promotes full-length SMN2 transcript formation, has significantly improved SMA outcomes. However, standardized in vitro procedures for evaluating nusinersen efficacy remain limited. This study aimed to optimize in vitro efficacy assessment of nusinersen across two human cellular models. Methods: Experiments were performed using HEK293 cells and the SMA patient-derived fibroblast line GM03813. Transfection conditions were optimized for each model. In HEK293 cells, several seeding densities were evaluated for nucleofection, while in GM03813 fibroblasts, multiple transfection reagents and protocols were tested. Nusinersen activity was quantified at the transcript and protein levels, and dose-response curves were generated to determine EC50 values. Results: In HEK293 cells, a higher seeding density (1 × 106 cells) yielded the most efficient nucleofection. In GM03813 fibroblasts, Lipofectamine 3000 outperformed the other transfection reagents tested. Nusinersen exhibited dose-dependent effects in both models. The EC50 for transcript induction in HEK293 cells was 293 nM, whereas in GM03813 fibroblasts the EC50 was 10 nM, demonstrating substantial model-dependent differences in response. Conclusions: This study establishes optimized conditions for in vitro efficacy assessment of nusinersen in HEK293 and GM03813 cellular models. These protocols provide a robust and reproducible framework for evaluating nusinersen and can be readily applied to other antisense oligonucleotides designed to correct SMN2 splicing.

Mesenchymal stromal cells (MSCs) are central to regenerative medicine and advanced therapies. However, the absence of consensus on reporting kinetic parameters, such as population doubling level (PDL), population doubling time (PDT), and the reliance on passage number alone obscures biological age and manufacturing history, and limits correlation of potency with expansion dynamics. Here, we clarify the distinctions among passages, PDL, PDT, and replication rate; we synthesize evidence that identical passage numbers can conceal multifold differences in cumulative doublings, with downstream effects on transcriptomic stability, and immunomodulatory performance. We further highlight culture determinants, oxygen tension, seeding density, media formulation, surface/bioreactor systems, and early niche mimetic stimuli, that shape proliferative kinetics and cellular aging trajectories in WJ-MSCs. Critically, we propose extracellular vesicles (EVs) as sensitive functional readouts of bioprocess stress and expansion history: EV quantity can increase while functional bioactivity declines, and EV miRNA cargo captures cell state programs not evident from minimal identity markers. To address these gaps, we recommend a reporting framework that incorporates: (1) culture conditions, (2) passage number and PDL at harvest, and (3) functional consequences of expansion. Adopting kinetic metrics beyond passage number will harmonize data capture and enable pooled analyses, accelerating clinical translation while safeguarding patient outcomes.

Mitochondrial dysfunction and chromatin dysregulation are interconnected contributors to neuronal vulnerability in Alzheimer's disease (AD), yet the molecular mechanisms linking these processes remain poorly understood. CHCHD10, a mitochondrial intermembrane space protein, has been implicated in neurodegenerative disorders, but its role in AD has not been defined. Here, we identify CHCHD10 as a previously unrecognized modulator of neuronal epigenomic stability in AD. Using direct fibroblast-to-neuron reprogramming, which preserves patient-specific epigenetic signatures, we show that AD neurons recapitulate genome-wide hypomethylation patterns observed in postmortem AD cortex. CHCHD10 expression is significantly reduced in AD neurons and across multiple human brain datasets, including single-cell and bulk RNA sequencing, proteomics, and human cortical tissue analyses. Restoration of CHCHD10 in AD neurons reduces amyloid-β and insoluble tau accumulation while reversing AD-associated differentially methylated regions across CpG islands, promoters, and regulatory elements. CHCHD10-responsive methylation changes overlap with those observed in human AD brain regions and colocalize with significant AD loci and cortex-specific eQTL loci, including MAPT and ABCA7. Finally, we identify KATNAL2 as a CHCHD10-responsive effector whose loss enhances tau phosphorylation and seeding, whereas its restoration mitigates tau pathology. Together, these findings support a CHCHD10-associated neuroprotective pathway linking mitochondrial dysfunction, epigenomic instability, and tau pathology in AD.

Porous hydrogels are critical for tissue engineering and regenerative medicine, as they mimic the native extracellular matrix to support cell infiltration and mass transport. A common strategy for engineering pore structures involves the incorporation and subsequent removal of sacrificial porogen templates (e.g., crystals or microspheres). Although this approach offers excellent control over pore architecture, it often suffers from complex procedures and biosafety concerns arising from incomplete template removal. In this work, we present a simple, biocompatible, and versatile templating approach. By systematically investigating the coacervation parameters, we produced gelatin microspheres (GSs) with tunable diameters from 7 µm to 300 µm via a green, instrument-free, and scalable process. Using GSs of 20-160 µm as porogens, we obtained alginate hydrogels with adjustable viscoelasticity, stiffness, and pore sizes. We then validated two cell-loading strategies for bulk porous alginate hydrogels using immortalized human T (Jurkat) cells: (i) post-seeding into pre-formed pores supported high-density, long-term, and organized cell aggregates with >90% viability; (ii) in situ encapsulation (prior to pore formation) yielded >80% viability and preserved the cluster-forming growth characteristics of Jurkat cells. Moreover, composites of smaller GSs (7-20 µm) with alginate could be syringe-extruded into stable, sub-millimeter porous filaments, demonstrating the potential for 3D printing. Collectively, this work provides a promising platform for three-dimensional culture of immune cells.

Periprosthetic joint infection (PJI) is one of the most serious complications of total joint arthroplasty (TJA), carrying both severe functional consequences and substantial economic costs. PJIs most commonly result from microbial contamination at the time of surgery or during the early postoperative period, while haematogenous seeding from distant infection sources accounts for a smaller proportion of cases. Patient-related factors, including diabetes mellitus, immunosuppression, obesity, malnutrition and smoking, as well as procedure-related factors such as prolonged operative time and wound complications, further increase risk. Among potential sources of haematogenous spread, the oral cavity has long been considered a possible source of infection, leading to routine dental clearance and prophylactic antibiotic use before dental procedures in patients with joint prostheses. However, contemporary epidemiological evidence challenges the causal role of dental-procedure-related bacteraemia in PJI. This review synthesises contemporary literature on the burden of PJI, the prevalence of dental pathology among arthroplasty patients, the microbiology of oral-associated infections and shifts in guideline recommendations. Across systemic reviews, cohort studies and registry data - dental associated PJIs are exceedingly rare. Prophylactic antibiotics provide no measurable protection and nearly half of the implicated organisms are resistant to amoxicillin. The evidence supports prioritising lifelong oral hygiene and targeted management of systemic comorbidities as more effective strategies for risk reduction. Selective dental intervention in high-risk patients, rather than universal clearance or prophylaxis, aligns with antibiotic stewardship and equitable access to surgery. Historical literature has not demonstrated a consistent causal relationship between dental pathology and PJI, highlighting the need for well-designed prospective studies. Accordingly, practice has shifted away from blanket antibiotic prophylaxis towards individualised, risk-based decision making.

Cardiovascular disease remains the predominant cause of mortality globally. Vascular grafts serve as substitutes to restore blood flow by replacing or bypassing diseased or occluded blood vessels. The application of small-diameter vascular grafts (SDVGs, diameter ≤ 6 mm) is limited due to their poor long-term patency primarily caused by acute thrombosis and chronic neointimal hyperplasia after implantation. Seeding cells to simulate the structure and function of the native endothelium represents an effective strategy to improve long-term patency. However, the effect of endothelialization has been constrained by the limited availability and functional heterogeneity of endogenous endothelial progenitor cells (EPCs), as well as the lack of biomimetic cellular organization on graft surfaces. In this study, we differentiated human induced pluripotent stem cells (hiPSCs) into EPCs (hiPSC-EPCs), which exhibited excellent pro-angiogenic function and the potential to inhibit inflammation. To promote biomimetic endothelialization, hiPSC-EPCs were seeded onto aligned electrospun nanofibers (ESNFs) fabricated for SDVGs. The aligned ESNFs guided hiPSC-EPCs to adopt a spindle-shaped morphology and an oriented cellular arrangement. Furthermore, the aligned hiPSC-EPC layer reduced platelet adhesion and inflammatory factor expression, while promoting the expression of endothelial functional markers in vitro. Our study combines hiPSC-EPCs with aligned ESNFs to closely simulate the structure and function of the native endothelium, thereby demonstrating a promising strategy to develop biomimetic SDVGs for clinical applications.

Recombinant AAV vectors are among the most extensively studied vectors for viral gene delivery due to their unique safety profile and their ability to mediate efficient, long-term transgene expression by persisting episomally in the nucleus. These properties make AAV vectors promising not only for the treatment of monogenic diseases but also for tissue regenerative applications. In the context of critical-sized bone defects, current gold-standard treatments are often associated with severe side effects, highlighting the need for alternative therapy strategies. In this study, we therefore developed a gene-activated matrix (GAM) for localized AAV-mediated gene delivery for potential applications in bone regeneration, establishing a workflow that is straightforward and transferable to other therapeutic settings. Following an initial screening of AAV serotypes and transgene DNA formats, reporter gene-expressing AAV2 vectors were associated with chitosan-based scaffolds containing varying amounts of β-tricalcium phosphate (β-TCP). Analysis of AAV release revealed that incorporation of β-TCP significantly reduced AAV release from 15.7% to approximately 6.6%. Furthermore, seeding of primary ovine mesenchymal stromal cells (oMSC) onto AAV-loaded scaffolds demonstrated efficient in situ delivery and expression of the osteogenic and angiogenic growth factors BMP-2 and VEGF in vitro. To further enhance AAV-mediated gene delivery, a panel of poloxamers was screened, leading to the identification of novel transduction enhancer AAVBlast. AAVBlast stabilized AAV particles and increased their bioavailability, resulting in significantly elevated intracellular AAV DNA levels, enhanced transgene mRNA expression, and increased protein production across multiple cell types. Modular application of AAVBlast onto the GAM significantly enhanced transduction of scaffold-released AAV particles but did not significantly affect transduction of oMSC by GAM-retained AAV vectors. In summary, this study demonstrates the identification of novel transduction enhancer AAVBlast and the successful development of a gene-activated matrix enabling efficient, localized AAV-mediated gene delivery in vitro, providing a promising platform for future GAM applications.

Ungernia victoris and U. sewerzowii (Amaryllidaceae J.St.-Hil.) are rare medicinal species of Central Asia known as sources of biologically active alkaloids, including galantamine. In this study, antioxidant activity was comparatively evaluated across different types of plant material, including natural populations, botanical garden specimens, in vitro regenerants, callus cultures, in vitro obtained bulbs, and seeds. Micropropagation systems based on direct and indirect organogenesis were developed using Murasige and Skoog and Vollosovich et al. media with various plant growth regulator combinations. Antioxidant activity was determined with the use of DPPH and ABTS assays and expressed as IC50 values. Significant variability was observed depending on population origin, type of biological material, and in vitro cultivation conditions. U. sewerzowii demonstrated higher antioxidant activity than U. victoris in natural populations. The highest activity was recorded in callus cultures, whereas in vitro-derived bulbs showed relatively low activity. A strong positive correlation between DPPH and ABTS assays confirmed the reliability of the results and indicated the contribution of multiple types of secondary metabolites. These findings highlight the potential of Ungernia callus cultures as a promising biotechnological platform for the production of antioxidant-active compounds and support sustainable utilization strategies.

The presence of fungi in seeds has a negative impact on germination and quality, which eventually leads to fungal deterioration and economic losses. This research inquired into the variety of fungi that were isolated from Foeniculum vulgare seeds (70 samples) collected from different regions of India, namely Delhi, Pondicherry, Rajasthan, Uttar Pradesh, and West Bengal. In order to identify the fungal genera, protocols outlined by the International Seed Testing Association (ISTA) (1980) procedures were adopted, and mycological media like, Potato Dextrose Agar (PDA) was used for both qualitative and quantitative analyses. Twenty-five fungal species that belonging to 11 different genera were isolated. Aspergillus niger, A. flavus, Mucor sp., Mycelia sterilia, Penicillium sp. and Yeast, were the most common types of fungi that were encountered in the present study. This is the first report of five Aspergillus spp., being isolated and documented from F. vulgare. One of the most significant contaminants, A. flavus was frequently isolated, with a toxigenic potential. The other types of fungi that are being reported, include Fusarium oxysporum, F. verticillioides, Alternaria alternata, Curvularia sp., and Trichothecium sp. Macromorphological and Micromorphological analyses were carried out after incubation at 25 °C for 7 days. Based on the outcome of the present mycological study, F. vulgare seeds contain an array of fungal contaminants, which have a potential effect on the health and viability of the seeds. In addition, they pose a serious threat to consumers because of their ability to produce mycotoxins.

Anxiety disorders are characterized by dysregulation of monoaminergic signaling and remain a significant therapeutic challenge due to limitations associated with current pharmacological treatments. In this context, the essential oil of Morisonia flexuosa (Capparaceae) seeds was chemically characterized and evaluated for anxiolytic-like activity in adult zebrafish. Chemical profiling by GC-MS and GC-FID revealed a predominance of isothiocyanates, particularly butyl isothiocyanate (42.60%) and isobutyl isothiocyanate (42.28%). Acute toxicity assessment demonstrated no lethality at the tested doses. Behavioral analyses showed a significant increase in light preference in the light/dark paradigm, with moderate locomotor reduction insufficient to account for the behavioral shift solely by sedation. Pharmacological antagonism assays indicated that the anxiolytic-like effect was predominantly mediated by 5-HT1 and 5-HT2A/2C receptors. Chemometric analyses (PCA, HCA, and heatmap) revealed statistical association between compound abundance and behavioral endpoints, supporting the contribution of major isothiocyanates within the tested model. Notably, the strongest behavioral response was observed at the lowest concentration, suggesting an ideal effective concentration range. Collectively, these findings provide the first evidence that an isothiocyanate-rich essential oil from M. flexuosa exerts serotonergic-involved anxiolytic-like effects in zebrafish and supports further mechanistic investigation of its neuropharmacological potential.

The endometrium exhibits the ability to regenerate without scarring after menstruation or parturition. Postpartum uterine repair occurs in a unique environment, shaped by peripartum immune responses. However, the mechanisms leading to healing or excessive fibrosis remain poorly understood. This study aimed to evaluate the drastic postpartum changes and the involvement of the immune milieu in the process of full wound regeneration. Publicly available scRNA-seq data of human myometrium from non-pregnant or postpartum day 7 individuals were analyzed. Allogenically mated and virgin C57/BL6 female mice served as models for human postpartum healing. Uterine tissues were analyzed by flow cytometry, immunohistochemistry, immunofluorescence, and quantitative real-time polymerase chain reaction. Human myometrial scRNA-seq revealed a sustained upregulation of immune responses in a context of degrowth of muscle components by postpartum day 7. As shown in mice, the size of the myometrium rapidly retracts. Particularly, the gradual resorption of the main site of leukocyte infiltration during pregnancy and postpartum, the mesometrial triangle, grants restoration of myometrial layers, dispersed during pregnancy to accommodate the leucocyte aggregates and growing conceptus. This region, encircling the lesions left by placental detachment, abundantly recruits macrophages that remain CD206neg. This inflammatory reaction coincides with elevated levels of Tnf, Il2, and Ifng, neovascularization, and cell death, and is balanced by increased Il10 levels. CD8+ T cells also seeded the mesometrial triangle, along with myofibroblasts surrounding the placental detachment clot until healing. This immune response was confined to the mesometrial area by a collagen-rich capsule. In turn, neighboring myometrial macrophages, largely expressing CD206, expanded during early remodeling to retract at homeostasis. In healthy postpartum uterus tissue, we identified the myometrium, endometrium, and mesometrial triangle as distinct anatomical compartments undergoing remodeling, each with unique yet interconnected processes. Within these, the distribution of F4/80+CD206+ and F4/80+CD206neg macrophage populations corresponded to different levels of tissue disruption. Inflammation, highly restricted by a collagen capsule and enhanced IL10, resolved to endorse myometrium integrity. Whether these mechanisms safeguard the myometrium from excessive immune responses, often linked to fibrosis, requires further empirical investigation.

Seed maturation is a critical developmental phase during which seeds acquire traits essential for nutritional value, desiccation tolerance, and long-term survival. Abscisic acid (ABA) signalling is a key regulator of this process, coordinating gene expression programs underlying the acquisition of seed quality traits. However, the molecular regulation of many of these traits remains poorly understood. To address this, we performed a comprehensive analysis of seed maturation in Arabidopsis thaliana, combining physiological and transcriptomic approaches across wild-type plants and mutants affected in ABA biosynthesis, signalling, and catabolism. We generated a high-resolution transcriptome dataset covering seed development from 12 days after pollination to the dry seed stage in wild-type and ten mutant lines. In parallel, we characterized the temporal acquisition of multiple seed traits, including germination capacity, dormancy, chlorophyll fluorescence, longevity and desiccation tolerance. Integration of these datasets using weighted gene co-expression network analysis (WGCNA) identified gene modules associated with specific trait acquisition patterns. This approach enabled the identification of coordinated transcriptional programs linked to distinct seed quality traits, extending beyond individual gene-level analyses. Notably, modules associated with desiccation tolerance and longevity were enriched for genes involved in stress responses and ABA-regulated pathways, highlighting the complex and multifactorial regulation of these traits. This study provides a comprehensive physiological and transcriptomic framework for understanding seed maturation and the acquisition of key seed quality traits in Arabidopsis thaliana. By linking gene expression dynamics to trait development, our work offers new insights into the regulatory networks underlying seed resilience and storage capacity. The dataset is made accessible through SeedMatExplorer (https://www.bioinformatics.nl/SeedMatExplorer), an open-access web platform that enables interactive exploration and supports hypothesis generation. Together, this resource represents a valuable tool for advancing research on seed biology and improving seed performance in agricultural contexts.

Polymeric procyanidins (PPCs) constitute the major fraction of procyanidins, but they have poor bioactivity. The purpose of this study is to clarify the composition and content of PPCs from Vitis amurensis Rupr. (Vitaceae) seeds before and after depolymerization, thereby providing a theoretical basis for activity evaluation and application of proanthocyanidins (PCs). PPCs extracted from V. amurensis seeds were depolymerized by catechin-assisted sulfitation. The compositions and contents of PCs before and after depolymerization were qualitatively and quantitatively analyzed by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), high-performance liquid chromatography (HPLC), and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Results showed that twenty-eight components were identified (7 monomers, 13 dimers, 5 trimers, 1 tetramer and 2 unknowns). Before depolymerization, tetrameric and higher polymers dominated, accounting for 58.81% of the relative content. After depolymerization, these high-molecular-weight compounds declined to <1% or became undetectable, while monomers and dimers (with minor trimers) surged to 42.89%. Among them, the relative content of two monomers and three dimers, catechin, epicatechin gallate and procyanidin B1-B3, increased by 37.00, 3.75, 10.98, 3.72 and 9.74 times, respectively. In conclusion, the method utilizing catechin-assisted sulfitation effectively depolymerizes PPCs from V. amurensis seeds into oligomeric components such as monomers and dimers.

The evolutionary arms race between bacteria and phages has driven the diversification of prokaryotic antiviral defense mechanisms, with nucleic acid degradation emerging as a central strategy. Here, we investigate a Higher Eukaryotes and Prokaryotes Nucleotide-binding (HEPN) domain-containing defense system from Escherichia coli that mediates RNase-dependent abortive infection. In contrast to canonical immune systems, where oligomerization triggers signaling, this system adopts a dodecameric autoinhibited architecture, with RNase activity unleashed upon oligomer dissociation. This activation mechanism is reminiscent of the dispersal of dandelion seeds, and we therefore term this defense system "Dandelion." We further identify the phage single-stranded DNA-binding (SSB) protein as a trigger for the Dandelion system, and phylogenetic analysis of SSB proteins uncovers the specificity underlying phage resistance. Our findings reveal a counterintuitive paradigm in bacterial immunity-&#x200c;oligomer disassembly as an activation switch, which challenges the long-standing dogma that protein oligomerization activates immune signaling.

Structural health monitoring (SHM) of fibre-reinforced composites requires a health indicator that is monotonically non-decreasing under the standard SHM assumption that no self-healing or maintenance-induced restoration event is active, derived from heterogeneous sliding-window observations of acoustic emission, strain, and fibre Bragg grating channels, with only the failure timestamp available per specimen. Conventional endpoint-supervised regressors attain high rank correlation with normalised life but produce jagged, non-monotone trajectories of limited engineering value. A method named SAMS-Net (Smoothness-Anchored Monotone Neural Differential Equation Network) is developed, in which a neural differential equation backbone is anchored by a two-level Pool-Adjacent-Violators (PAV) projection. A within-window projection is applied during training with a straight-through gradient, and an across-window projection is applied at inference, yielding a globally non-decreasing health indicator. A smoothness-stratified two-phase training schedule first trains on specimens whose per-specimen median local-smoothness coefficient exceeds 0.5, then fine-tunes on the full set. Across the present seventeen-specimen open-hole carbon-fibre dataset spanning two stress levels and six leave-one-specimen-out and cross-condition scenarios, SAMS-Net wins on every scenario on the canonical Prognostics and Health Management (PHM) Composite of monotonicity, trendability, and robustness, with margins of 0.22 to 0.48 against the strongest baseline, reproducible across three random seeds. Ablation reveals that the operative mechanism is the two-level PAV projection rather than the stochastic differential equation (SDE) inductive bias. A new control experiment in which the across-window PAV projection is applied at inference to the strongest baselines confirms that the projection accounts for a substantial share of the SAMS-Net margin, while the within-window training-time projection and a globally consistent prognosability metric retain a SAMS-Net advantage. Cross-site or cross-material transferability remains to be established in future work.

In this study, the effects of five bean seeds (Vigna radiata, Pisum sativum, Vigna unguiculata, Vicia faba, and Glycine max) on the development, survival, oviposition, and digestive enzyme activities of C. chinensis were investigated. The duration of C. chinensis development from egg to adult was shortest when reared on V. radiata (26.97 d) and longest on G. max (30.92 d) (p < 0.01). Similarly, the emergence rate of C. chinensis was highest on V. radiata (66.11%) and lowest on G. max (33.89%) (p < 0.01). Significant differences in the fecundity of C. chinensis were also detected, with the highest values on V. radiata (63.36 eggs/female), followed by P. sativum (54.27 eggs/female), V. unguiculata (51.30 eggs/female), V. faba (42.47 eggs/female), and G. max (37.29 eggs/female) (p < 0.01). For an equal initial insect number, after continuous rearing for 30, 60, or 90 d, there were significantly more offspring on V. radiata than on the other species, with the fewest offspring detected on G. max (p < 0.01). Although CL activity (p = 0.33) in C. chinensis did not differ significantly among the different legume species, activities of PEP (p < 0.01) and &#x3b1;-AMS (p < 0.01) in C. chinensis were all highest on V. radiata and lowest on G. max, and activities of LPS (p < 0.01) were highest on G. max and lowest on V. radiata. The differences in population dynamics associated with different bean seeds may be related to the activity of particular digestive enzymes, which play important roles in insect nutrient metabolism and growth. Our results indicated that V. radiata is the most suitable host diet, whereas G. max is the least suitable for the development of C. chinensis.

The European legislation sets the maximum residue levels for glyphosate in sesame seeds at 0.1 mg/kg (EU Regulation n. 293/2013) and for glufosinate and N-Acetyl-glufosinate expressed as glufosinate at 0.03 mg/kg (EU Regulation n. 2016/1002). The present work describes a rapid methodology to determine glyphosate, glufosinate and its metabolite and N-Acetyl-glufosinate in sesame seeds by LC/MS/MS. The method was studied in the framework of EU proficiency tests on sesame seeds. The analytical method was developed using methanol acidified with formic acid (1%, v/v) extraction with an isotope internal standard, followed by LC/MS/MS detection. The recoveries were performed in the range of 0.05-0.5 mg/kg for glyphosate and 0.02-0.2 mg/kg for glufosinate and N-Acetyl-glufosinate. All the recovery values were between 70 and 114%, which is the acceptable interval according to SANTE/11312/2021; the relative standard deviation (%RSD) values met the requirement of <20%. Linearity for each substance in solvent and matrix was studied, and the response was linear with R2 > 0.999. We considered precision, matrix effect, LOD and LOQ in the validation. All the parameters were in agreement with the acceptability criteria of the document SANTE/11312/2021. The method was considered suitable for the determination of the studied substances on sesame seeds.

The edible seeds of pumpkin plants (genus Cucurbita) are becoming increasingly appreciated as functional foods for their nutritional benefits, medicinal properties, and bioactive compounds, including lipids, proteins, and antioxidants. Particularly, the naked seeds of Cucurbita pepo var. styriaca have proved to yield both an edible oil showing anti-inflammatory properties in treating skin disorders and hydro-alcoholic extracts effective in inhibiting the growth of cancer cells. In this study, a detailed and extensive analysis of the eco-friendly alcoholic extract of the seeds of this variety was accomplished by using LC-HRMSMS techniques, with the main aim to broaden the knowledge on bioactive lipids other than the already reported fatty acids. The obtained results highlighted the occurrence of numerous compounds belonging to different classes of polar and neutral lipids, such as phospholipids, sphingolipids, glycolipids, acylglycerols, and oxylipins. Noteworthily, a significant presence of Cer-(EO)LCBs, i.e., Cer-EOS-type ceramides with different long chain base (LCB) and fatty acid composition, was detected, representing a real novelty for pumpkin. Additionally, a good number of multiflorane-type triterpenoids were detected, only some of which were previously reported in this plant. These findings highlight the nutraceutical value of these edible seeds.