Search Results (11,120)

Background/Objectives: The multi-attribute method (MAM) has found diverse use in the analytical characterization of therapeutic protein products during their development and production. As the MAM matures it has the potential to enter quality control (QC) laboratories, consolidating and replacing many less informative chromatographic techniques; however, this requires an appropriate risk assessment and understanding of method capability. Methods: A validated MAM approach was used to quantify product quality attributes (PQAs) using three different mass spectrometers across two laboratories; the results were compared to conventional hydrophilic interaction chromatography–fluorescence detection (HILIC-FLD) and cation exchange chromatography–ultraviolet (CEX-UV) techniques. Results: Stressed, long-term, and accelerated stability studies were performed, and their effects on glycosylation, deamidation, oxidation and N- and C-termini were quantified. Conclusions: Overall, the inter-instrument inter-laboratory data provided here showed important considerations for transferring methods between laboratories and establishing the correlation between the MAM and conventional data, elements which are necessary to transition the MAM to the QC environment and ultimately achieving the goal of replacing orthogonal QC methods. Full article

The reduced form of nicotinamide adenine dinucleotide phosphate (NADPH) is a primary electron donor for both antioxidant enzymes, such as glutathione reductase, and pro-oxidant enzymes, such as NADPH oxidases that produce reactive oxygen species (ROS) and nitric oxide synthases that generate nitric oxide which act as signaling molecules. Monitoring NADPH levels, NADPH/NADP⁺ ratio, and especially distinguishing from NADH, provides vital information about cellular redox status, energy generation, survival, lineage specification, and death pathway selection. NADPH detection is key to understanding metabolic reprogramming in cancer, aging, and cardiovascular, hormonal, neurodegenerative, and autoimmune diseases. Liquid chromatography combined with mass spectrometry (LC-MS) is crucial for NADPH detection in redox signaling because it offers the high sensitivity, specificity, and comprehensive profiling needed to quantify this vital but labile redox cofactor in complex biological samples. Using hepatoma cell lines, liver tissues, and primary hepatocytes from mice lacking transaldolase or nicotinamide nucleotide transhydrogenase, or having lupus, this study demonstrates that accurate measurement of NADPH depends on its preservation in reduced form which can be optimally achieved by extraction of metabolites in alkaline solution, such as 0.1 M potassium hydroxide (KOH) in comparison to 80% methanol (MeOH) alone or 40:40:20 methanol/acetonitrile/formic acid solution. While KOH extraction coupled with hydrophilic interaction liquid chromatography (HILIC) and mass spectrometry most reliably detects NADPH, NADP, NADH, NAD, polyamines, and polyols, MeOH extraction is best suited for detection of glutathione and overall discrimination between complex metabolite extracts. This study therefore supports performing parallel KOH and MeOH extractions to enable comprehensive metabolomic analysis of redox signaling. Full article

Multi-epitopes of FMDV can be used to develop a novel vaccine. Determining how to screen naturally presenting epitope peptides derived from FMDV is crucial for advancing progress in this area. In this study, a transient expression plasmid named pEGFP-N1-VP1 was transfected into Porcine Kidney Epithelial cells 15 (PK15). The positive cells that stably expressed the O-VP1 gene of FMDV were screened with gradient concentrations of G418 (Geneticin). The constructed pEGFP-N1-VP1/PK15 cell line was eluted by pH 3.3 phosphate buffer to isolate the eluted peptides, followed by desalting, liquid chromatography–tandem mass spectrometry (LC–MS/MS), a flow cytometric analysis of SLA-I expression, and an ELISA detection of SLA-I bound peptides. It was demonstrated that a PK15 cell line stably expressing the VP1 gene was initially screened out at 500 μg/mL of G418, followed by culturing at 300 μg/mL. The O-VP1 expression was identified using an image analysis system, RT-PCR, and Western blot analysis. Thirty-seven peptides derived from O-VP1 were eluted from the constructed cell line. The flow cytometric analysis and ELISA detection results showed that the eluted peptides were associated with SLA-I and bound. This is the first known study to construct a cell line for screening naturally presenting antigenic peptides derived from the O serotype of FMDV. Full article

Natamycin is widely used in other countries for the postharvest treatment of agricultural commodities to prevent fungal growth. However, since no MRL has been set in Korea, natamycin residues are regulated under the Positive List System (PLS) with a uniform limit of 0.01 mg/kg, requiring the development of highly sensitive and reliable analytical methods. In this study, a QuEChERS-based analytical method was developed and validated for the quantification of natamycin in five agricultural commodities—soybean, mandarin, hulled rice, green pepper, and potato—using liquid chromatography–tandem mass spectrometry (LC-MS/MS). Extraction using methanol with 3 g of MgSO₄ resulted in high recoveries without crystallization, and clean-up with MgSO₄ and C18 effectively reduced matrix interferences blow <50%. Natamycin was detected in all five matrices at 6.8 min without any interfering peaks. The MLOQ was determined at 0.01 mg/kg for all five matrices. The mean recoveries (82.2–115.4%) and %CV values (1.1–4.6%) values were within the acceptance criteria defined by the CODEX guidelines. Matrix effects were classified as “soft” for mandarin (|ME| < 20%) and “medium” for soybean, hulled rice, green pepper, and potato (20% ≤ |ME| < 50%). The analytical method for natamycin was validated as suitable for regulatory safety monitoring under the Korean PLS. Full article

Cicadas are a traditional food in China. In this study, we performed multielement analysis on cicadas using Inductively Coupled Plasma–Mass Spectrometry (ICP-MS). In total, 40 cicada samples were collected in Hebei Province (China). Comparing the results of mushroom and vegetable samples selected as the standard foods among the eight food categories, which had the highest correlation with other food categories, there are 12 elements (Al, Mn, Zn, Cu, Cd, Ba, Se, V, As, Li, Pb and Co) in cicadas both higher than mushroom and vegetable samples, which a total of 25 elements were detected. The Principal Component Analysis (PCA), cluster analysis, and correlation analysis were used in the data analysis. HQ (Hazard quotient) value for Cd, As, Pb, and Al is 0.281 in males and is 0.324 in females. Concentrations of essential trace elements (Zn, Cu, and Se) in cicadas ranged from 1.7 to 101.9 times higher than those in mushrooms or vegetables. We analyzed in detail the two perspectives: one is the risk of potentially toxic elements, and the other is the enrichment of essential trace elements. In this study, we reveal that cicadas show a low risk of potentially toxic elements and high concentrations of essential trace elements, making them an edible insect. Full article

Insulators on power lines require regular maintenance by operators in high-altitude hazardous environments, and the emergence of aerial manipulators provides an efficient and safe support for this scenario. In this study, a lightweight telescopic aerial manipulator system is developed, which can realize the installation and retrieval of insulator inspection robots on power lines. The aerial manipulator has three degrees of freedom, including two telescopic scissor mechanisms and one pitch rotation mechanism. Multiple types of cameras and sensors are specifically configured in the structure, and the total mass of the structure is 2.2 kg. Next, the kinematic model, dynamic model, and instantaneous contact force model of the designed aerial manipulator are derived. Then, the hybrid position/force control strategy of the aerial manipulator and the visual detection and estimation algorithm are designed to complete the operation or complete the task. Finally, the lifting external load test, grasp and installation operation test, as well as outdoor flight operation test are carried out. The test results not only quantitatively evaluate the effectiveness of the structural design and control design of the system but also verify that the aerial manipulator can complete the accurate automatic grasp and installation operation of the 3.6 kg target device in outdoor flight. Full article

Ovarian cancer’s high mortality is primarily due to late-stage diagnosis, underscoring the critical need for improved early detection tools. This study develops and validates explainable artificial intelligence (XAI) models to discriminate malignant from benign ovarian masses using readily available demographic and laboratory data. A dataset of 309 patients (140 malignant, 169 benign) with 47 clinical parameters was analyzed. The Boruta algorithm selected 19 significant features, including tumor markers (CA125, HE4, CEA, CA19-9, AFP), hematological indices, liver function tests, and electrolytes. Five ensemble machine learning algorithms were optimized and evaluated using repeated stratified 5-fold cross-validation. The Gradient Boosting model achieved the highest performance with 88.99% (±3.2%) accuracy, 0.934 AUC-ROC, and 0.782 Matthews correlation coefficient. SHAP analysis identified HE4, CEA, globulin, CA125, and age as the most globally important features. Unlike black-box approaches, our XAI framework provides clinically interpretable decision pathways through LIME and SHAP visualizations, revealing how feature values push predictions toward malignancy or benignity. Partial dependence plots illustrated non-linear risk relationships, such as a sharp increase in malignancy probability with CA125 > 35 U/mL. This explainable approach demonstrates that ensemble models can achieve high diagnostic accuracy using routine lab data alone, performing comparably to established clinical indices while ensuring transparency and clinical plausibility. The integration of state-of-the-art XAI techniques highlights established biomarkers and reveals potential novel contributors like inflammatory and hepatic indices, offering a pragmatic, scalable triage tool to augment existing diagnostic pathways, particularly in resource-constrained settings. Full article

Background/Objectives: Therapeutic drug monitoring (TDM) of antipsychotic medications plays an important role in optimizing treatment efficacy, reducing adverse effects, and supporting adherence. While Ultra-High Performance Liquid Chromatography–Tandem Mass Spectrometry (UHPLC–MS/MS) has long been the gold standard for antipsychotic quantification, recent advances in automated platforms and microsampling raise questions about its current clinical practicality. This systematic review evaluated the clinical applicability and analytical performance of UHPLC-based methods for monitoring antipsychotic drugs, focusing on precision, recovery, matrix effects, and suitability across various biological matrices. Methods: A systematic search of PubMed, Scopus, and Web of Science was conducted for studies published between 2013 and 2024 involving UHPLC-based quantification of antipsychotics in clinical samples from adult patients. Data on analytical parameters, sample matrices, and study characteristics were extracted. A custom quality checklist was used to assess methodological rigor. In addition to qualitative synthesis, non-traditional quantitative approaches were applied, including descriptive aggregation of recovery, matrix effects, and precision across studies, as well as correlation analyses to explore relationships among performance parameters. Results: Twelve studies were included, spanning a range of typical and atypical antipsychotics and metabolites. Plasma and serum demonstrated the highest analytical reliability (recovery >90%, minimal matrix effects), while dried blood spots (DBSs), whole blood, and oral fluid showed greater variability. Clinically, UHPLC–MS/MS enabled more accurate dose adjustments and identification of non-adherence, outperforming immunoassays in sensitivity, specificity, and metabolite detection. Microsampling methods showed promise for outpatient and decentralized care but require further clinical validation. Conclusions: UHPLC–MS/MS remains the most robust and reliable method for TDM of antipsychotics, especially when quantification of active metabolites is required. While logistical barriers remain, technological advances may enhance feasibility and support broader integration into routine psychiatric care. Full article

In vitro fertilization (IVF) exposes embryos to environmental stressors that can disrupt early development and confer long-term health risks, though the mechanisms remain poorly understood. Here, we tested the hypothesis that reducing incubation temperature during the first zygotic cleavage would promote long-term developmental stability in IVF-conceived offspring. Using a mouse model, we compared the long-term effects of standard (37 °C) versus reduced (35 °C) IVF culture temperature on energy balance, circadian rhythms, sleep architecture, and brain histone modifications. Although offspring from both IVF groups exhibited increased body mass without notable effects on glucose metabolism, significant disruptions in circadian rhythms and sleep–wake patterns were detected. The 37 °C group exhibited altered amplitudes in oxygen consumption rhythms and respiratory exchange ratios, as well as pronounced alterations in sleep–wake patterns, including reduced sleep duration and increased nighttime activity. The 35 °C group displayed intermediate phenotypes, substantiating the importance of optimizing embryo incubation parameters. These metabolic and behavioral changes were paralleled by altered histone modifications in the cerebral cortex of IVF offspring, suggesting an epigenetic basis for circadian misalignment. Our results identify disrupted circadian rhythm and sleep architecture as a novel mechanism contributing to metabolic dysfunction in IVF-conceived offspring. The partial mitigation of these effects through reduced culture temperature underscores the importance of optimizing IVF protocols to minimize long-term epigenetic and metabolic risks. Full article

Capsaicinoids (CPCs) are regarded as a typical marker of waste oil, which has emerged as a serious food safety issue in developing countries, necessitating the development of rapid, sensitive, and specific detection methods. In this study, a novel hapten was synthesized to generate a high-affinity monoclonal antibody (mAb) targeting CPCs. Subsequently, aggregation-induced emission fluorescent microspheres (AIEFMs), known for their superior fluorescence intensity, were utilized as an enhanced probe to develop a lateral flow immunoassay (LFIA) based on mAb 8B4 for CPC detection. For comparison, a traditional gold nanoparticle (AuNP)-LFIA was also constructed using the corresponding mAb. The AIEFM-LFIA demonstrated a limit of detection (LOD) of 0.33 µg/kg for CPCs in edible oil samples, which is 4.21 times lower than the LOD of 1.39 µg/kg achieved by the AuNP-LFIA. And the assay effectively identified three additional CPCs, with LODs ranging from 0.26 to 0.99 µg/kg, while exhibiting minimal cross-reactivity with CPC analogs, indicating high specificity. The recovery rates of the AIEFM-LFIA in oil samples ranged from 75.0% to 106.0%, with coefficients of variation ≤ 8.3%, exhibiting excellent accuracy and precision. Furthermore, the results of the AIEFM-LFIA demonstrated a strong degree of correlation with liquid chromatography–tandem mass spectrometry, with a correlation coefficient (R²) of 0.978. Consequently, the developed AIEFM-LFIA shows significant promise as a rapid, sensitive, specific, and reliable method for detecting CPCs in oil samples. Full article

Background: Fabry disease (FD) is an X-linked lysosomal storage disorder caused by GLA mutations, leading to deficient α-galactosidase A (α-Gal A) activity and progressive glycosphingolipid accumulation. While α-Gal A activity is the diagnostic gold standard, its sensitivity is reduced in late-onset or heterozygous patients. Conventional biomarkers such as lyso-Gb3 provide only limited insight into disease progression and therapeutic response. Exosomes, as stable carriers of disease-specific proteins, may offer complementary biomarkers for early detection and longitudinal monitoring. Methods: Twenty-one pediatric FD patients with confirmed GLA mutations were enrolled. Clinical, enzymatic, renal, and cardiac parameters were assessed. Plasma-derived exosomes were characterized by transmission electron microscopy and proteomic profiling. Differentially expressed proteins were identified using mass spectrometry, analyzed using GO/KEGG enrichment, and validated using RT-PCR, ELISA, and immunofluorescence in patient samples and Gla^−/− mice. Results: Male patients showed markedly reduced α-Gal A activity and elevated lyso-Gb3 compared with females. Although overt renal and cardiac dysfunction was uncommon, several patients exhibited early abnormalities such as proteinuria, an elevated LVMI, or increased cTnI levels. Proteomic analysis identified 2553 proteins, of which 188 were differentially expressed. Fibrosis- and inflammation-related proteins, including THBS1 and CFHR5, were upregulated, while protective factors such as APM1, SERPINA10, and CAB39 were downregulated. IGFBP3 was also elevated and closely linked to tissue remodeling. Enriched pathways were involved in PPAR/AMPK signaling, lipid metabolism, and complement activation. Conclusions: Exosomal proteomic profiling revealed early molecular signatures of cardiorenal involvement in pediatric FD. Key proteins such as THBS1, CFHR5, IGFBP3, APM1, and CAB39 show strong potential as biomarkers for risk stratification, disease monitoring, and therapeutic evaluation. Full article

Background: Steamed Panax notoginseng saponins (SPNSs) can alleviate cyclophosphamide-induced anemia. However, the hepatointestinal effects of SPNSs and their role in ameliorating cyclophosphamide-induced anemia remain unexplored. Objective: To elucidate the hepatointestinal effects of SPNSs and their role in ameliorating cyclophosphamide-induced anemia. Methods: Blood samples were collected and analyzed on days 7 and 14. Liver tissues and small intestinal villi structures were observed via HE staining. Liver and colon content metabolites were detected by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Liver proteins were analyzed by using an Orbitrap Astral mass spectrometer. Colon content microbiota composition was assessed via metagenomics. Signaling pathway protein expression was analyzed via Western blotting (WB). Results: SPNSs significantly increased the red blood cell (RBC) count and hemoglobin (HGB) level by day 14 and alleviated hepatointestinal damage. Hepatic metabolomics revealed: the most abundant metabolites were fatty acids and stachyose on day 7 and amino acid and arachidonic acid derivatives on day 14. KEGG analysis implicated cAMP signaling. Proteomics revealed upregulated immune-related proteins and enhanced PI3K pathway activity (WB-validated). Colon content metabolomics showed increased daidzein, 3-(2,5-dimethoxyphenyl) propanoic acid, γ-CEHC, and adenosine in SPNS groups on day 14. Metagenomics indicated differential abundances of Heminiphilus faecis, Phocaeicola sartorii, and s-bacterium_J10.2018 on day 14. Multiomics integration demonstrated significant correlations between hepatic metabolites, hematopoietic proteins, colon content metabolites, and probiotic bacteria. Conclusions: SPNS alleviates cyclophosphamide-induced hepato-intestinal injury in anemic mice by modulating the gut microbiota and enhancing hepato-intestinal immune defense. Additionally, SPNSs ameliorate anemia in cyclophosphamide-treated mice by activating the cAMP/PI3K/AKT pathway, promoting hepatocyte proliferation, and increasing hematopoietic protein expression. Full article

Echocardiography remains a vital part of the initial assessment and monitoring of oncological patients. It allows for proper treatment selection but can also reveal life-threatening complications, including impaired left ventricular function or thromboembolism. It can rarely detect intracardiac masses that require further investigation. In the presented case, a 51-year-old female patient with left-sided breast cancer, who had undergone neoadjuvant chemotherapy, was hospitalised due to a right atrial mass identified via routine transthoracic echocardiography (TTE). Initial anticoagulation therapy showed no clinical improvement. Follow-up TTE revealed a 12 × 19 mm hyperechogenic, mobile mass in the right atrium (RA). Computed tomography angiography (CTA) ruled out pulmonary embolism and revealed that the mass was located close to the tip of the vascular access port. Transoesophageal echocardiography showed that the lesion was not connected to the vascular port. Based on location and mobility, the lesion was most consistent with a cardiac myxoma. After the Heart Team made a decision, endovascular intervention using a vacuum-assisted device was performed without complications. Histopathological examination excluded thrombosis and myxoma, revealing a fibro-inflammatory lesion. A multimodality approach is necessary to assess RA masses. However, even an extensive evaluation could be misleading, so treatment options should always be subject to the Heart Team’s decision. Full article

Background: Impaired balance is one of the most common and functionally limiting problems in children with cerebral palsy (CP), significantly affecting their motor abilities and quality of life. Although force platforms are considered the gold standard for evaluating postural stability, they are often costly, non-portable, and require specialized laboratory environments, limiting their accessibility in routine clinical settings. Objective: This study aimed to develop a novel software program based on image processing techniques to assess static balance in children with CP and to evaluate its validity against traditional force platform measurements. Methods: A total of 83 children aged 5–15 years (63 with CP, GMFCS levels I–II; 20 healthy controls) participated. Static balance was assessed under four different standing conditions using both a force platform and a newly developed video-based software tool. The software utilized the frame difference method to detect center of mass movements, and parameters such as velocity and total displacement were calculated. Correlation analyses were conducted between the image processing and force platform data. Results: The software demonstrated moderate to strong positive correlations with force platform parameters in the majority of test conditions, particularly when participants stood with eyes open. In more challenging balance scenarios (e.g., eyes closed, feet together), correlations were weaker but still significant. Conclusions: The findings suggest that this image-based software is a valid, low-cost, and portable alternative for static balance assessment in children with CP. It has the potential for use in diverse clinical or home settings, supporting individualized rehabilitation strategies. Full article

Pancreaticobiliary endoscopy, encompassing endoscopic ultrasound (EUS), endoscopic retrograde cholangiopancreatography (ERCP), and digital single-operator cholangioscopy (DSOC), is essential for diagnosing and managing pancreatic and biliary diseases. However, these procedures are limited by operator dependency, variable diagnostic accuracy, and technical complexity. Artificial intelligence (AI), particularly through machine learning (ML) and deep learning (DL), has emerged as a promising tool to address these challenges. Early studies show that AI can enhance lesion detection, improve differentiation of pancreatic masses, classify cystic lesions, and aid in diagnosing malignant biliary strictures. AI has also been used to predict post-ERCP pancreatitis risk and reduce radiation exposure during ERCP. Despite this promise, current AI models are largely experimental—limited by small, single-center datasets, lack of external validation, and no FDA-approved systems for these indications. Major barriers include inconsistent data acquisition, limited interoperability across hardware platforms, and integration into real-time workflows. Future progress depends on multicenter data sharing, standardized imaging protocols, interpretable AI design, and regulatory pathways for model deployment and updates. AI can be developed as a valuable partner to endoscopists, enhancing diagnostic accuracy, reducing complications, and supporting more efficient, personalized care in pancreaticobiliary endoscopy. Full article