Search Results (1,235)

Cow-based calf rearing (dam- or foster cow-based) is gaining popularity as a relatively natural husbandry practice. However, direct udder contact by calves creates a bidirectional transmission route for pathogens. The objective of this cross-sectional study was to investigate the prevalence and concordance of mastitis-associated pathogens in the milk of cows and the saliva of corresponding calves at the time of weaning. On 15 organic dairy farms, milk samples from 269 cows and saliva samples from 403 calves were collected at weaning. Pathogen differentiation was performed via Matrix-assisted laser desorption time-of-flight mass spectrometry (MALDI-TOF), and strain typing was additionally conducted via randomly amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR). Dominant pathogens in milk were Non-aureus-Staphylococci (NaS)/Mammaliicoccus spp. and Corynebacterium spp., and in calf saliva, NaS/Mammaliicoccus spp. (primarily M. sciuri, S. xylosus). There were a total of 12 strain concordances, exclusively in foster cow–calf pairs (Fisher’s exact test, p = 0.012). The highest observed/expected ratio was found for P. multocida (21.1). For S. aureus, strain matches were found in 3 foster cow–calf pairs across 2 farms. Direct strain concordance was generally observed rather rarely. The observed strain concordances suggest transmission between cows and calves, particularly for S. aureus and P. multocida. NaS and Mammaliicoccus spp. concordances are likely attributable to a common environmental source. Full article

Background: The m⁷G cap structure, which mimics the natural cap of eukaryotic mRNA, is a critical determinant of mRNA vaccine efficacy, safety, and stability. However, its precise quantification remains challenging due to complex impurity profiles and the high physicochemical similarity between the target cap and related impurities. Although liquid chromatography mass spectrometry (LC-MS) is widely employed for this purpose, current methodologies still face significant limitations, including labor-intensive sample preparation, low analytical throughput, poor reproducibility in quantifying low-level impurities, and a lack of universally applicable strategies across diverse mRNA vaccine platforms. Methods: We systematically optimized sample preparation and LC-MS detection workflows. RNase H-mediated cleavage was compared with DNAzymes, guide DNA probes were rationally designed, and thermostable RNase H was introduced for one-step denaturation and cleavage. To establish an accurate, efficient, and universal sample preparation workflow. Chromatographic conditions were optimized using an ion-pairing reagent system to suppress ESI-MS metal adducts. Eliminating sample purification improves recovery, reduces manual handling errors, and boosts assay efficiency. Results: Through optimally designed guide DNA probes, RNase H cleavage specificity reached ≥98% with high cleavage efficiency, offering higher efficiency than DNAzyme. Furthermore, the incorporation of thermostable RNase H enabled a single-step workflow combining high-temperature denaturation and site-specific cleavage, substantially streamlining sample preparation. On the chromatographic side, optimization of the ion-pairing reagent system effectively suppressed metal adduct formation in electrospray ionization mass spectrometry (ESI-MS). This advancement enabled direct injection of the 5′ cap fragments without purification, achieving high-recovery quantification while demonstrating broad compatibility across mainstream LC-MS platforms. The optimized assay reduces the total analytical workflow from 4~6 h to under 1.5 h. Conclusions: Combining high accuracy, robustness, and broad platform compatibility, this method offers a universal, high-throughput analytical solution for mRNA vaccine quality control and continuous process development. Full article

Background: Phosphatidylcholine (PC) is a major phospholipid that contributes to intestinal barrier protection and is essential for hepatic secretion of lipids and bile acids. Because inflammatory bowel disease (IBD) and primary sclerosing cholangitis (PSC) are closely linked, we hypothesized that individual serum PC species would reflect disease activity. We therefore investigated whether serum PC profiling could identify clinically useful biomarkers across the gut–liver axis. Methods: Serum concentrations of 21 PC species were quantified by direct flow injection high-resolution mass spectrometry in 16 healthy controls, 57 patients with IBD, and 20 patients with PSC. Results: In IBD, multiple serum PC species were inversely associated with inflammatory activity, showing negative correlations with serum C-reactive protein and fecal calprotectin. Patients with fecal calprotectin concentrations above the diagnostic cut-off of 120 µg/g had lower levels of PC 34:3, 36:1, 36:2, 36:3, 36:4, 36:5, 38:3, 38:4, 38:5, 38:7, 40:5, and 40:6, as well as lower total PC. In contrast, in PSC, PC 30:0, 32:0, 32:1, and 34:1 were increased compared with IBD and correlated positively with gamma-glutamyltransferase and alkaline phosphatase. Furthermore, these shorter-chain PC species as well as PC 36:1 were markedly elevated in PSC with advanced liver fibrosis compared with PSC without fibrosis. Conclusions: Serum PC species show a reciprocal disease-associated pattern in IBD and PSC. In IBD, lower concentrations of predominantly unsaturated PC species are associated with active intestinal inflammation, whereas in PSC, higher concentrations of shorter-chain PC species are associated with cholestatic injury and advanced liver fibrosis. IBD and PSC exhibit opposing serum PC signatures, suggesting that dysregulated PC metabolism is a pathophysiological feature of intestinal inflammation and PSC-associated liver fibrosis. Full article

Protein-based biosensors require controlled and site-selective functionalization strategies to enable stable and oriented immobilization without compromising protein structure and signal transduction efficiency. We evaluated a chemoselective linchpin-directed modification (LDM) approach targeting Lys–His pairs as a tool for site-specific labeling of the model fluorescent biosensor green fluorescent protein (GFP). LDM molecules with variable spacer lengths were prepared, and a structure-guided computational workflow was implemented to map Lys–His distances on the protein and identify candidate modification sites. Experimental validation by UV-Vis spectroscopy and mass spectrometry demonstrated efficient conjugation and a final degree of labeling close to unity, consistent with single-site modification, with all LDM molecules selectively targeting the same histidine residue (His181), independently of spacer length. Structural analysis revealed that this residue is located within an accessible internal cavity that favors productive interactions with the reactive group. Importantly, the modification preserves GFP fluorescence and pH response, confirming retention of sensing functionality. These results demonstrate that LDM enables selective modification not only of surface residues, but also of structurally guided, non-surface residues. This approach provides the proof of concept of a new, promising strategy for the controlled functionalization and immobilization of protein-based biosensors. Full article

Processed meat products represent a major challenge for proteomic species identification due to extensive thermal treatment and protein structural changes. In this study, species-specific peptides in pork, chicken, and bovine meat products were identified using a directed fragmentation-assisted de novo sequencing workflow that combines 4-formylbenzene-1,3-disulfonic acid (FBDA) peptide derivatization, dual-polarity data-independent mass spectrometry (DIA-MS), and Protein Acrobat de novo sequencing software. Comparative analysis of non-fractionated and strong cation exchange (SCX)-fractionated pork luncheon samples improved peptide and protein identification after fractionation, with 312 peptides and 115 protein groups detected exclusively in fractionated samples. Species-specific peptides were predominantly assigned to conserved muscle-related proteins, including myosin, troponin, and tropomyosin, while sequence variability enabled reliable species discrimination despite protein conservation across species. To evaluate applicability for food fraud detection, mixed meat samples containing 10% chicken in pork or bovine matrices were analyzed, reflecting potential economically motivated adulteration through substitution with lower-cost meat components. Several chicken-specific peptides remained detectable in both mixtures, demonstrating robustness of the FBDA-assisted peptide sequencing combined with SCX fractionation and DIA-MS for detection of adulteration in complex processed food matrices. These findings establish a mass spectrometry-driven orthogonal method to ELISA testing for fast, reliable and accurate metaproteome analysis of highly processed food. Full article

Background: Inborn errors of metabolism (IEMs) represent a diverse group of genetic disorders affecting biochemical pathways. Despite advances in diagnostic technologies, comprehensive understanding of their historical evolution, classification systems, and diagnostic approaches remains fragmented. Objectives: This systematic review and meta-analysis aimed to synthesize evidence on the historical development, classification frameworks, and diagnostic modalities for IEMs, diagnostic accuracy, and prevalence estimates, providing a comprehensive resource for clinicians and researchers. Methods: Following PRISMA 2020 guidelines, we conducted a systematic search of seven electronic databases (PubMed/MEDLINE, Embase, Scopus, Web of Science, Google Scholar, SciSpace and ArXiv) from January 2000 to March 2026. Studies addressing historical perspectives, classification systems, or diagnostic approaches for IEMs were included. Two independent reviewers performed screening, data extraction, and quality assessment. Meta-analyses were conducted using random-effects models for diagnostic accuracy and prevalence estimates. Results: From 1342 identified records, 54 studies met the inclusion criteria, encompassing 8,234,567 individuals across 35 countries. Historical analysis revealed 16 major milestones from Garrod’s 1902 “chemical individuality” concept to the current AI-powered diagnostics. Four major classification systems were identified: pathophysiological (intoxication, energy deficiency, complex molecule disorders), biochemical pathway (amino acid, organic acid, urea cycle, carbohydrate, fatty acid oxidation, mitochondrial, peroxisomal, lysosomal disorders), organelle-based, and the integrated Society for the Study of Inborn Errors of Metabolism (SSIEM) nosology. Meta-analysis demonstrated high diagnostic performance of tandem mass spectrometry (MS/MS) with a pooled sensitivity of 99.1% (95% CI: 98.6–99.5) and specificity of 99.8% (95% CI: 99.7–99.9%). The pooled global prevalence of IEMs was 50.9 per 100,000 live births (95% CI 45.2–56.8). Next-generation sequencing achieved a diagnostic yield of 42.8% (95% CI: 38.2–47.5%) in suspected cases. Emerging AI-powered diagnostic tools demonstrated high discrimination performance with area under the curve (AUC) values exceeding 0.95 for specific IEM, though external validation remains limited. Newborn screening expanded from single-disease to comprehensive panels detecting over 50 disorders. Conclusions: This comprehensive review demonstrates that IEMs have evolved from rare curiosities to systematically diagnosable conditions through technological advances. Integration of metabolomics, genomics, proteomics and artificial intelligence promises further diagnostic improvements. Standardized classification systems and evidence-based diagnostic algorithms are essential for optimal patient care. Future directions include artificial intelligence-enhanced diagnostics, expanded screening, and personalized medicine approaches. Full article

Background/Objectives: Direct-fed microbials (DFMs) are widely used in dairy calves to improve gut health and mitigate neonatal disorders, yet their systemic metabolic effects remain poorly defined. This study evaluated the impact of DFM supplementation on the plasma metabolome of pre-weaned dairy calves using untargeted liquid chromatography–mass spectrometry (LC–MS). Methods: Eighty-six Holstein bull calves (2 to 5 days old) were assigned to one of four treatments in a 2 × 2 factorial randomized complete block design: Lactobacillus plantarum in starter (CLP), a culture mix of Bifidobacterium animalis and Lactobacillus animalis in milk replacer (BBCM), and a combination of both (CMLP), or no supplementation (CON). Blood samples collected on days 0 and 56 were subjected to metabolomic profiling, and metabolites were annotated using Human Metabolome Database and Kyoto Encyclopedia of Genes and Genomes databases. Results: A total of 231 plasma metabolites were detected. Compared with CON, 24 metabolites were differentially abundant in DFM-treated calves (fold change ≥ 1.2 or ≤ 0.83; p ≤ 0.05). Supplemented calves exhibited increased abundances of ketone functional groups, aldehydes and amino acid-related metabolites. Metabolite set enrichment analysis identified 11 significantly enriched pathways. Branched-chain amino acid degradation pathways (valine, leucine, and isoleucine) were enriched in CLP and CMLP calves, whereas carbohydrate metabolism pathways, including pentose and glucuronate interconversions, were enriched in the CLP and BBCM groups. Conclusions: These findings demonstrate that DFM supplementation modulates systemic metabolism in dairy calves, particularly pathways involved in amino acid and carbohydrate utilization, suggesting enhanced metabolic efficiency during early life. Full article

As global ecosystems and food systems face unprecedented anthropogenic and climatic challenges, there is a demand for an integrated understanding of biological systems. Proteomics has emerged as a definitive approach offering a direct view of the molecular phenotype, yet it is traditionally separated into plant and animal disciplines. With recent advances in mass spectrometry (MS) and bioinformatics tools, this prospective review proposes that combining a One Health proteomics approach with deep-learning data analysis can revolutionize global food security, animal productivity, and ecosystem health by uncovering proteoform signatures that drive resilience across life. The potential of a unified One Health proteomic framework, highlighting major developments, including 4D proteomics, Data-Independent Acquisition (DIA), and single-cell resolution, and emphasizes their capacity to resolve the complex proteoform landscape across kingdoms. Review emphasizes the applications of proteogenomics as a cross-disciplinary tool to improve genome annotations, explain evolutionary differences, discover biomarkers in animals and resolve complex signaling networks in plants under stress. Nevertheless, contemporary proteogenomics methods still show limitations in their ability to comprehensively resolve proteoforms due to the fact that the use of peptide-based approaches makes it difficult to fully appreciate the post-translational modifications specific to each protein isoform. We show that One Health proteomics will provide a transformative roadmap for deciphering the functional proteoform signatures that underpin resilience across the tree of life. Full article

The Nuri deposit is currently the only Cu-W-Mo deposit in the Gangdese metallogenic belt, Tibet, China, that contains large-scale tonnages for both Cu and WO₃ resources, accompanied by a medium-scale Mo resources. Previous studies have suggested the potential presence of concealed porphyry-type orebodies at depth, yet effective exploration tools for verifying this hypothesis remain lacking. In this study, microscopic identification, electron probe microanalysis (EPMA), and laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) were integrated to investigate the mineral chemical characteristics of chlorite from the Nuri deposit. The aim was to evaluate the effectiveness of chlorite geochemistry as an exploration vector for predicting deep concealed porphyry orebodies and to establish corresponding exploration indicators. Chlorite in the deposit can be genetically classified into metasomatic (Chl-I) and hydrothermal (Chl-II) types. Both types are Mg-rich varieties, indicating formation under conditions of low oxygen fugacity and low pH. With decreasing vertical distance to the orebody and toward the southeast direction of the exploration section, the contents of Ti (10–950 ppm) and V (50–820 ppm), as well as the Ti/Sr, Ti/Mn, Ti/Li, and V/Li ratios, progressively increase. In contrast, the concentrations of Li (36–390 ppm), Mn (1270–6730 ppm), Sr (1–510 ppm), and Zn (110–1100 ppm) systematically decrease. These systematic compositional variations demonstrate that chlorite geochemistry is an effective exploration tool in the Nuri mining area and suggest the presence of a concealed mineralization center or porphyry orebody beneath the interval from ZK4501 to ZK4502. Full article

Human saliva is a heterogeneous bodily fluid with a complex composition, which contains antibodies, proteins, and viruses, making it applicable in clinical diagnosis. There are several advantages of the analysis of saliva samples over other biofluids, including a non-invasive and simple collection procedure for extraoral detection. Biomarker or pathogen detection in saliva can be performed with various methods: mass spectrometry, PCR, ELISA, electrochemical, and optical methods such as fluorescence, SPR, and SERS. The early detection of cancer and other disease biomarkers, as well as infectious agents, can be crucial for effective treatment and minimization of mortality from those diseases. The following paper reviews extraoral detection techniques to identify the most sensitive methods for diagnosing early and asymptomatic patients. The LODs collected and tabulated from 149 analytical papers, alongside the sensitivity, specificity, and sometimes the area under the curve (AUC) tabulated from 118 clinical studies, have all become parameters for the comparative quantitative analysis. Based on the limited but substantial number of analytical studies on the detection of cortisol in saliva (29), the electrochemical platforms demonstrated the highest sensitivity, with a geometric mean LOD of 11 pM. Within these methods, voltametric ones showed the best performance with 6 pM geometric mean LOD. Electrochemical techniques are then followed by immunoassay- and mass spectrometry-based platforms, with corresponding geometric average LOD values of 39.1 and 171 pM, respectively. However, clinical outcomes are at least as meaningful as LOD values. In terms of clinical analysis, ELISA and direct-SERS outperformed other methods, achieving balanced accuracy of approximately 87% and AUC values of 0.96 for direct SERS and 0.86 for ELISA. MS and PCR followed closely, with balanced accuracies around 84%. While the direct SERS is not yet widespread in clinical applications, its potential can be forged if the standardization issue is addressed. Full article

Medulloblastoma is one of the most prevalent pediatric brain tumors. Currently, existing therapies for this devastating type of cancer can only prolong survival time with severe side-effects and relapse. These therapies are not curative for almost a third of treated patients, while most survivors are condemned to a poor quality of life. The addition of immune checkpoint inhibitors (ICIs) to immune therapy has given some hope to those suffering from this type of cancer. Although ICIs provide a valuable contribution to immunotherapy, the exploitation of immune checkpoint inhibition within existing therapeutic strategies to cure Medulloblastoma remains understudied. However, the identification of the main molecular subgroups of medulloblastoma is considered one of the success stories of oncology. This advancement in molecular profiling of MB paved the way to subgroup-directed clinical trials, which may lead to efficacious immune-targeted therapy. However, this relatively new development is still hampered by a substantial biological heterogeneity of the disease and the absence of a full understanding of the various mechanisms behind its resistance to existing therapeutic modalities. The inclusion of chimeric antigen receptor (CAR) T and CAR NK cell therapy within various therapeutic strategies and ongoing clinical trials has given fresh hope those suffering from this fatal disease. However, ongoing clinical trials suggest that this highly promising therapy can be impaired by a number of serious limitations, including cytokine release syndrome, Graft-versus-host disease, the scarcity of target antigens, and severe adverse events. Some of the ongoing clinical trials also suggest that CAR NK is less prone to some of these limitations. This review also highlights the contribution of mass spectrometry-based proteomics, and the increasing role of liquid biopsy rather than tissue biopsy. Full article

Mass spectrometry-based lipidomics has created new opportunities to investigate the role of lipids in coffee quality formation and stability across the production chain. Coffee lipids contribute to flavor precursor formation, aroma release, mouthfeel, and storage behavior, but their molecular remodeling during maturation, processing, roasting, and storage remains insufficiently integrated. This review summarizes recent progress in lipidomics methodologies relevant to coffee research, with emphasis on sample preparation, mass spectrometry platforms, data analysis, and the strengths and limitations of current lipid annotation strategies. It further examines how lipid profiles change during bean maturation, how they differ among coffee species and varieties, and how they are reshaped by postharvest processing, roasting, and storage. However, it is important to note that most of these associations are currently correlational rather than causal; direct evidence linking specific lipid species to particular sensory attributes remains limited. Existing studies suggest that lipid composition, rather than total lipid content alone, is more informative for understanding coffee quality differences and for identifying candidate markers associated with origin, processing method, roasting degree, and storage conditions. In particular, alterations in glycerolipids, glycerophospholipids, fatty acids, diterpenes, and other minor lipid constituents are increasingly associated with lipid oxidation, thermal degradation, and flavor-related transformations in coffee. However, current evidence is still limited by incomplete structural annotation, isomeric ambiguity, platform dependence, and the frequent gap between statistical discrimination and mechanistic validation. Future work integrating high-resolution mass spectrometry, ion mobility, targeted quantification, stable isotope tracing, sensory analysis, and multi-omics approaches will be essential to improve marker reliability and to clarify the functional roles of coffee lipids. Overall, lipidomics provides a promising framework for linking molecular composition with coffee quality control, traceability, and process optimization, although substantial work is still needed to establish mechanistic links to flavor formation. Full article

Background and Objectives: Temporomandibular disorders (TMDs) encompass a broad spectrum of functional and structural abnormalities of the temporomandibular joint (TMJ). Conventional diagnostic tools, although essential, often fail to capture the underlying biochemical mechanisms driving disease progression. Synovial fluid (SF), by virtue of its direct proximity to intra-articular tissues, represents an accessible biological matrix for identifying molecular signatures of inflammation, cartilage degradation, lubrication failure, oxidative stress, and angiogenic activation. The objective of this review is to synthesize current evidence on SF proteomics in TMD and evaluate its potential translational value in precision medicine. Materials and Methods: A narrative review of the literature was conducted on PubMed to identify human studies focused on SF proteomic and biochemical biomarkers in TMD. Eligible studies included original research articles assessing SF composition in relation to specific TMJ pathologies, diagnostic categories, or clinical phenotypes. Extracted data included study design, sample characteristics, analytic methodology, biomarkers investigated, and key findings. Google Gemini (Google LLC, Mountain View, CA, USA) was used as an AI-assisted tool to support language editing and manuscript writing during the preparation of this article. The use of this tool was limited to linguistic refinement; all scientific content, data interpretation, and conclusions were formulated and verified by the authors. Results: Across the analyzed studies, TMD phenotypes—particularly disc displacement with or without reduction (DDwR, DDwoR) and osteoarthritis (OA)—were characterized by consistent alterations in cytokines (IL-1β, IL-6, IL-8, TNF-α), extracellular matrix (ECM) components (aggrecan, glycosaminoglycans (GAGs), decorin, MMP-2, MMP-9), lubrication molecules (lubricin/PRG4), oxidative stress mediators (myeloperoxidase (MPO), nitric oxide (NO), glutathione peroxidase (GPX)), adipokines (chemerin, resistin, adiponectin), and angiogenic factors (vascular endothelial growth factor (VEGF), fibroblast growth factor-2 (FGF-2)). Recent liquid chromatography–tandem mass spectrometry (LC–MS/MS) analyses further revealed phenotype-specific protein clusters and pathways related to inflammation, ferroptosis, hypoxia signaling, and proteoglycan metabolism. Conclusions: Current evidence suggests that SF proteomics and multi-analyte biomarker profiling offer a promising, hypothesis-generating approach for understanding the biological mechanisms underlying TMD. The integration of proteomic, metabolic, and inflammatory markers holds future potential for diagnostic panel development; however, prospective clinical validation is still required before SF-based molecular profiling can be implemented as a precision medicine tool in TMJ disorders. Full article

The Tonglushan ore field is an important component of the polymetallic mineralization belt in the middle and lower reaches of the Yangtze River in China. The skarn-type Cu, Fe, Au, and Mo molybdenum deposits are mainly developed in the contact zone between the rock mass and the strata, as well as in the contact zone between residual and capturing bodies in the rock body. The distribution of ore bodies is controlled by faults and strata, but there is a lack of large-scale geophysical information on the contact relationship between the ore-forming geological body and the host rock and on the deep spatial morphology of the ore-forming structure and intrusion rock. The study uses the JS-WEM2 wide-field electromagnetic instrument and the RS230 spectrometer to conduct the ground frequency domain electromagnetic and radiometric spectrometry measurements on four profiles. The measurement results indicate that the fault distribution in the Tonglushan ore field is predominantly in the NW-trending and NE-trending directions. The NW-trending Tonglushan–Lijiashan fault (F2) is a steeply dipping fault; the NE-trending faults are minor, with steep dips, generally extending no deeper than −1000 m. The Tonglushan stock exhibits the northeastward uplift, characterized by southward overlap and southeastward dip. The deep resistivity is greater than 3000 Ω·m, while the resistivity below −1000 m is less than 2000 Ω·m due to the fault influence. The ore bodies are mainly distributed along the contact zones where variations in the occurrence of the rock intersect with the strata. On resistivity profiles, these zones show the gradient variation in resistivity and the distorted shape of the resistivity contour line. The radioactive element contents of wall rock above the ore bodies are characterized by high U, high Th, and low K. The Wide-Field Electromagnetic Method (WFEM) can effectively detect the distribution and morphology of rocks and faults, and combined with the radioactive characteristics of geological bodies, it can effectively identify concealed faults and the favorable mineralization target areas. Novelty: The study combines the WFEM with radiometric measurements to reduce uncertainty in exploration compared to using only one method. It improves the detection accuracy and target identification ability of deep hidden ore bodies, providing the new technical method for deep mineral exploration in complex structural areas. Full article

The lysosomal enzyme α-galactosidase A (AGAL) degrades globotriaosylceramide (Gb₃). While this enzymatic function in lysosomal metabolism is well characterized, interaction partners and alternative functions are unknown. This study aims to identify new potential AGAL-interacting proteins. AGAL was fused to the mutated biotin ligase BirA from E. coli (TurboID). Expression of the fusion protein was confirmed by Western blot and immunofluorescence, while enzymatic activity was verified by functional assays. In three experimental settings (AGAL wild-type (WT), AGAL missense variant (p.N215S), and the control cell line), TurboID-biotinylated proximal proteins were enriched by streptavidin pull-down and analyzed by mass spectrometry. Gene Ontology (GO) terms were subsequently evaluated to characterize biological functions and localizations of the identified proteins. Selected candidates were co-immunoprecipitated with AGAL to confirm direct interactions. The AGAL-TurboID fusion protein was successfully expressed in AB8/13 podocytes. Immunofluorescence and enzyme activity assays confirmed the presence and functionality of the fusion protein. Subsequent functional analysis (GO term analysis) showed enrichment of driver terms, including extracellular matrix organization (ECM), multicellular organism development, and protein metabolic process, in the biological process category. The identified top-hit proteins were predominantly involved in the organization of ECM, cell proliferation and cytokinesis, unfolded protein response during endoplasmic reticulum stress, and protein ubiquitination. Co-immunoprecipitation confirmed the interaction between AGAL and the candidate Galectin-3-binding protein (Gal-3BP). Our results suggest that AGAL may play a role in other pathways and/or the ECM organization beyond its lysosomal function. The confirmed interaction with Gal-3BP can now be functionally investigated in further studies. Full article