Search Results (1,526)

Introduction: Gastrointestinal stromal tumors (GISTs) are primarily driven by mutations in KIT (KIT proto-oncogene receptor tyrosine kinase) or PDGFRA (platelet-derived growth factor receptor alpha), but resistance to tyrosine kinase inhibitors (TKIs) such as imatinib remains a major clinical challenge. Alterations in fibroblast growth factor receptor 2 (FGFR2), although rare, are emerging as important contributors to tumor progression and drug resistance. This review evaluates the molecular mechanisms, expression profiles, detection methods, and therapeutic implications of FGFR2 in GIST. Methods: We searched PubMed, Web of Science, Google Scholar, and ClinicalTrials.gov for studies published between January 2010 and June 2025, using combinations of keywords related to FGFR2, gastrointestinal stromal tumor, resistance mechanisms, gene fusion, amplification, polymorphisms, and targeted therapy. Eligible studies were critically assessed to distinguish GIST-specific data from evidence extrapolated from other cancers. Results:FGFR2 is expressed in multiple normal tissues and at variable levels in mesenchymal-derived tumors, including GIST. Its alterations occur in approximately 1–2% of GIST cases, most commonly as gene fusions (e.g., FGFR2::TACC2, <1%) or amplifications (1–2%); point mutations and clinically significant polymorphisms are extremely rare. These alterations activate the MAPK/ERK and PI3K/AKT pathways, contribute to bypass signaling, and enhance DNA damage repair, thereby promoting TKI resistance. Beyond mutations, mechanisms such as amplification, ligand overexpression, and microenvironmental interactions also play roles. FGFR2 alterations appear mutually exclusive with KIT/PDGFRA mutations but occasional co-occurrence has been reported. Current clinical evidence is largely limited to small cohorts, basket trials, or case reports. Conclusions:FGFR2 is an emerging oncogenic driver and biomarker of resistance in a rare subset of GISTs. Although direct evidence remains limited, particularly regarding DNA repair and polymorphisms, FGFR2-targeted therapies (e.g., erdafitinib, pemigatinib) show potential, especially in combination with TKIs or DNA-damaging agents. Future research should prioritize GIST-specific clinical trials, the development of FGFR2-driven models, and standardized molecular diagnostics to validate FGFR2 as a therapeutic target. Full article

Legionella pneumophila (L. pneumophila) infection is characterized by a wide spectrum of manifestations, from influenza-like illness to life-threatening atypical pneumonia with multiorgan failure. The aim of our study was the assessment of in vitro and ex vivo neutrophil activation in L. pneumophila infections, as well as the role of neutrophils’ peptides such as LL-37 in infection. The ability of neutrophils to form ex vivo extracellular traps (NETs) in response to bacterial infection was examined by immunofluorescence. In parallel, patients’ sera, as well as opsonized standard L. pneumophila strains, were used for in vitro activation of neutrophils from healthy individuals. The serum levels of interleukins were assessed using the LEGENDplexTM Multi-Analyte Flow Assay Kit. Furthermore, citrullinated cf-DNA as a marker of neutrophil extracellular traps (NETs) was detected in the serum of patients with acute infection. It was demonstrated that neutrophils released NETs in vitro and ex vivo upon L. pneumophila (interaction in an autophagy-independent manner. Notably, IL-1b was detected on NETs, but an antimicrobial peptide LL-37 was absent. The lack of antimicrobial activity failed to inhibit bacterial proliferation. In addition, in vitro and ex vivo NETs formation was observed during the Clarithromycin treatment. Those NETs were decorated with bioactive antimicrobial peptide LL-37, which inhibits bacterial proliferation. The findings provide evidence that neutrophils release NETs in vitro and ex vivo by expressing the IL1β protein in them. The lack of expression of the antimicrobial peptide LL-37 on the NETs demonstrates the inability of the cells to inhibit proliferation, and consequently the elimination of L. pneumophila. Clarithromycin plays a dual role in the elimination. Full article

Complications during pregnancy and parturition can lead to foetal hypoxia, which may be responsible for cardiac ischemia and the subsequent release of troponin from cardiac muscles into the amniotic fluid (AF) and bloodstream. So far, cardiac troponin T (cTnT) has only been measured in the blood samples of adult dogs, while no data on its presence and relevance in AF are available. This study aimed to determine whether cTnT can be detected in canine AF collected at birth. Furthermore, a possible correlation between amniotic cTnT concentration and maternal and neonatal outcomes was explored. For this purpose, 40 AF samples were collected from 14 bitches at the time of delivery. A commercially available ELISA kit was used for the analysis of canine cTnT in biological fluids. Cardiac troponin T was detected in all amniotic specimens with concentrations ranging from 74.1 to 318 ng/L (191.6 ± 66.4 ng/L). The dams’ morphotype, age, and weight, as well as the type of parturition (elective vs. emergency C-section) and the expulsion time of puppies, were significantly associated with amniotic cTnT concentrations. Although amniotic cTnT warrants further investigation to fully understand its clinical role in canine neonatology, these results suggest a promising and valuable contribution. Full article

Crimean-Congo hemorrhagic fever virus (CCHFV) is a tick-borne zoonotic pathogen of growing public health concern in southeastern Europe. This study provides the first serological evidence of CCHFV circulation in Croatia, based on testing 1473 serum samples from farm and companion animals, including sheep, horses, cattle, goats, dogs, and cats. A total of 109 samples (7.4%) tested positive for CCHFV antibodies using a commercially available enzyme-linked immunosorbent assay (ELISA) kit. The highest seroprevalence was recorded in sheep (28.3%), followed by horses (4.3%) and a single cat (0.5%), with no antibodies detected in cattle, goats, or dogs. Almost all seropositive animals originated from coastal and subcoastal Croatia, where Hyalomma ticks are present. Only two seropositive cases were detected in continental areas. Sheep samples from several farms in Zadar County showed intra-farm seropositivity rates of up to 85.7%, suggesting localised virus circulation likely influenced by vector distribution and farm-level practices. No viral ribonucleic acid (RNA) was detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR), consistent with the transient nature of viremia in most animal hosts. These findings confirm the silent circulation of CCHFV in Croatia and reinforce the need for targeted, regionally adapted surveillance strategies that integrate multiple hosts and support early warning systems aligned with the One Health concept. Full article

Parkinson’s disease (PD), a progressive neurodegenerative disorder affecting over 10 million people worldwide, necessitates continuous symptom monitoring to optimize treatment and enhance quality of life. Effective communication between patients and healthcare providers (HCPs) is vital but often hindered by fragmented data and cognitive impairments. PARKA AI, a novel iOS application, leverages Apple Watch HealthKit data (e.g., tremor detection, mobility metrics, heart rate, and sleep patterns) and integrates it with self-reported logs (e.g., mood, medication adherence) to empower PD self-management and improve patient–HCP interactions. Employing a human-centered design approach, we developed a high-fidelity prototype using a large language model (LLM)— Google Gemini 1.5 Flash—to process and analyze self-reports and objective sensor-derived data from Apple Healthkit to generate patient-friendly summaries and concise HCP reports. PARKA AI provides accessible data visualizations, personalized self-management tools, and streamlined HCP reports to foster engagement and communication. This paper outlines the derived design requirements, prototype features, and illustrative use cases to show how LLMs can be used in digital health tools. Future work will focus on real-world usability testing to validate the application’s efficacy and accessibility. Full article

Brucellosis is a significant public health challenge in Kenya, particularly in pastoralist communities where the disease is endemic. Reliable and accurate point-of-care diagnostics are essential for timely case identification and effective disease management. The Febrile Brucella Agglutination Test (FBAT) is commonly used for diagnosis of brucellosis in Kenya, but concerns have been noted about its diagnostic accuracy, prompting an independent evaluation. The aim of this study was to compare the diagnostic performance of five FBAT kits with a commercial Enzyme-Linked Immunosorbent Assay (ELISA) as the reference standard, and to build local laboratory capacity for in-house kit validation for the Kajiado County laboratory staff. A total of 200 serum samples (100 ELISA-confirmed positives and 100 negatives) were tested using the FBAT kits. Each kit was evaluated for its ability to detect antibodies to both B. abortus and B. melitensis antigens. Diagnostic performance indicators, including sensitivity, specificity, and Cohen’s Kappa, were calculated, and McNemar’s test was applied to assess concordance with the ELISA results. Overall, none of the FBAT kits proved to have acceptable sensitivity and specificity compared to ELISA. We conclude that FBAT kits are not sufficient for the diagnosis of brucellosis and that alternative diagnostics should be considered. Full article

Early diagnosis by newborn screening (NBS) has contributed to improved outcomes in children with cystic fibrosis (CwCF). Georgia’s two-tiered algorithm consists of a fixed immunoreactive trypsinogen (IRT) cut-off followed by a 39-variant CFTR genetic panel. We conducted a retrospective review of CwCF born in Georgia from 2007 to 2022 to evaluate false negative NBS frequency. We characterized CwCF whose diagnosis was delayed beyond 28 days of age despite positive NBS. Six cases were detailed demonstrating the impact of missed and delayed diagnoses. We examined IRT trends from 2018 to 2022 and cut-off approaches. Missed case detection by expanded CFTR variant assays was assessed. Of 390 CwCF born in Georgia, 18 (4.6%) had false negative NBS—6 due to lack of CFTR variant detection and 12 due to low IRT values. Thirty children had delayed diagnosis, with the majority related to sweat testing. Minoritized children made up 19% of the population but 43% of missed and 44% of delayed diagnoses. Black and Hispanic infants had higher odds of missed or delayed diagnosis compared to non-Hispanic White infants (OR = 2.7, p = 0.027 and OR = 6.1, p < 0.001, respectively). Average IRT values varied across kits and were lower in warmer seasons. Expanded CFTR assays would reduce missed cases. Our results informed recommendations for improvement at multiple steps in the NBS process. Full article

Background: The 99th-percentile upper reference limit (99% URL) is essential for high-sensitivity cardiac troponin I (hs-cTnI) assays to diagnose acute coronary syndromes. We derived the gender-specific 99% URLs for the new Snibe hs-cTnI assay and verified its high-sensitivity performance. Methods: The Snibe hs-cTnI assay has a claimed limit of blank/detection/quantitation of 0.5/1.0/2.0 ng/L, a precision of 5.67% (@ 9.83 ng/L), 4.65% (@ 21.0 ng/L) and 3.77% (@ 174 ng/L), an analytical measuring range of 1.00–500,000 ng/L, and a claimed 99% URL of 20.1/11.8/17.5 ng/L (males/females/overall). Assay precision was evaluated using kit control materials. A total of 846 (M 444, F 402) anonymized leftover samples from healthy individuals were assessed for the derivation of 99% URLs. Results: The inter-assay CV was 6.2/4.4% (@ 10.1/20.6 ng/L). The 10/20% CV corresponded to a concentration of 5.2/3.0 ng/L. The assay recorded detectable hs-cTnI values in 64% of women and 70% of men. Hs-cTnI values were significantly lower in females than males (females vs. males median 2.6 vs. 3.2 ng/L, Mann–Whitney p = 0.005). The derived 99% URL for the population was 7.7 ng/L (90% CI 6.93–22.8 ng/L) for women and 13.7 ng/L (90% CI 10.6–41.4 ng/L) for men. When confined to subjects ≥40 years old only, males had a 99% URL of 14.8 ng/L (n = 412, 90% CI 10.6–41.4 ng/L) and 8.1 ng/L (n = 377, 90% CI 6.95–22.8 ng/L) for females, respectively. Conclusions: The Snibe hs-cTnI assay fulfills the requirements of a hs-cTn assay. Gender-specific 99% URLs are derived for this assay and they increased with age. Full article

Objectives: This study focuses on the regulatory mechanism of Schisandrin B (Sch B) on the lipid metabolism and apoptosis of AML-12 liver cells, with a particular emphasis on its potential therapeutic effect and mechanism of action in preventing and treating metabolic-associated fatty liver disease (MAFLD) by activating the PPARγ signaling pathway. Methods: An MAFLD cell model was established by inducing AML-12 cells with a mixture of oleic acid (OA) and palmitic acid (PA) (2:1). AML-12 cells were divided into a control group, a model group, and 20 μM and 40 μM Sch B groups. The cells were lysed and prepared into the cell suspension, then the cell suspension was centrifuged to obtain its supernatant, and the levels of total cholesterol (TC), triglycerides (TG), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) in the supernatant were detected according to the instructions of the kits. Effects of Sch B on the pathological changes of AML-12 cells were observed by Oil Red O staining. The key targets were screened through network pharmacology, and relevant targets were verified through molecular docking simulation. The activity of PPARγ was detected using a dual luciferase reporter plasmid, and the level of cell apoptosis was detected using the Annexin V-FITC/PI double staining method. The Western blot method was used to analyze the expression of genes related to lipid metabolism and apoptosis pathways. Results: Sch B could regulate lipid metabolism disorders in OA+PA-induced MAFLD cell model. The activation of PPARγ-PCK1/Aspase is a key step in the action of Sch B, which can effectively block fatty acid synthesis, improve fatty acid oxidation, and reduce lipid droplet aggregation in liver cells, thereby alleviating lipid metabolism abnormalities in the MAFLD cell model and inhibiting cell apoptosis. Conclusions: This finding may lay an important theoretical foundation and open a new research direction for the deep development and application of Schisandra chinensis. Full article

Background/Objectives: Adalimumab and Infliximab are biologics used to treat autoimmune diseases. Monitoring drug and anti-drug antibody (ADA) levels in patients helps optimize treatment. However, current quantitation methodologies for drug and total (free and drug-bound) ADAs often involve multi-step workflows. Automated systems can streamline the process. The i-Tracker chemiluminescent immunoassays (CLIA) are cartridge-based kits for quantifying serum levels of drugs such as Adalimumab, Infliximab, and associated ADAs. Herein, we aimed to establish performance characteristics of the i-Tracker Adalimumab, Infliximab, and total ADAs in serum on the random-access analyzer IDS-iSYS and to compare patient results with an electrochemiluminescent immunoassay (ECLIA)-based reference method. Methods: Remnant serum specimens, calibration material, or spiked serum were used to evaluate assay linearity, precision, functional sensitivity, and accuracy on the IDS-iSYS analyzer and to perform the method comparison. Results: The assays displayed linearity, accuracy, and up to 8% imprecision across clinically relevant analyte ranges. Compared to the reference method, the drug assays exhibited a strong linear fit (correlation coefficient > 0.95) with <±1.0 µg/mL mean bias. The total anti-Adalimumab assay demonstrated over 85% qualitative agreement. The total anti-Infliximab assay, however, showed higher detection rate of ADAs in Infliximab-treated patient specimens, yielding < 60% negative agreement with the reference method. Although i-Tracker total ADA assays exhibited drug sensitivity, they still detected ADAs in supratherapeutic drug concentrations. Conclusions: The i-Tracker assays demonstrated robust analytical performance, suggesting potential for clinical application. The method comparison underscored functional differences with the reference method, an important consideration when transitioning assay formats for monitoring Adalimumab- and Infliximab-treated patients. Full article

Introduction: Mn is a trace element essential for growth and development in organisms, and adequate Mn levels are crucial for maintaining normal liver function. This study aimed to investigate the effects of Mn deficiency on the liver and elucidate the underlying mechanisms using transcriptomics. Methods: Weanling mice were fed a Mn-deficient diet, and Mn chloride (MnCl₂) was administered intraperitoneally to correct the deficiency. Liver pathological changes were evaluated through histological examination. Liver function and key lipid metabolism markers were assessed using biochemical assays, while hepatic oxidative stress levels were measured via flow cytometry and biochemical kits. Alterations in inflammatory factors were detected using ELISA and qPCR. The mechanisms underlying Mn’s effects on liver function were further explored through Western blot, qPCR, and transcriptome sequencing. Results: Mn deficiency impaired liver morphology and structure. Serum levels of ALT, AST, and ALP were significantly elevated, while ALB decreased, confirming hepatic dysfunction. This dysfunction led to oxidative stress, characterized by increased hepatic ROS and MDA levels, alongside reduced Mn-SOD, GSH-Px, and T-AOC activities. Additionally, Mn deficiency elevated serum TG, TC, and LDL-C levels, indicating abnormal lipid metabolism. Hepatic pro-inflammatory factors (IL-6, IL-1β, and TNF-α) were significantly upregulated. Transcriptomic analysis revealed distinct gene expression patterns under different Mn conditions, with KEGG pathway analysis identifying the PPAR signaling pathway as a key regulatory target. Conclusions: Our findings suggest a potential pathogenic cascade in which manganese deficiency may initially induce hepatic oxidative stress, potentially leading to suppression of the PPAR signaling pathway. This inhibition of PPARα/γ could subsequently orchestrate downstream manifestations of aberrant lipid metabolism and inflammatory responses. Thus, the PPAR signaling pathway is proposed as a plausible central hub for translating oxidative damage into metabolic and inflammatory dysfunction in the manganese-deficient liver. Full article

Background/Objectives: Most cervical intraepithelial neoplasias (CINs) regress over time. Diagnostic screenings are limited and cannot identify the disease trend, which leads to the risk of overtreatment. Reliable methods are needed to preselect patients who will probably progress. The diagnostic GynTect^® assay offers sensitive and specific CIN identification from cervical scrapes, measuring the methylation of six marker genes. We studied the main marker (ZNF671) methylation on formalin-fixed paraffin-embedded (FFPE) material to determine if the kit provides prognostic information too. Methods: We tested 289 FFPE samples from 139 patients with varying CIN grades and disease trends, including regressive, persistent, progressive, and recurrent disease. Additionally, we correlated age and human papillomavirus (HPV) status with the results. Results: Although there are differences between FFPE material and cervical scrapes, we achieved a similar increase in ZNF671 methylation with increasing neoplasia grade (dysplasia-free: 0%, CIN 1: 8.20%, CIN 2: 26.73%, CIN 3: 32.43%, carcinoma: 100%). In addition, ZNF671 is more likely to detect recurring (27.12% of positives) and progressive (59.32% of positives) neoplasia. Patients with regressive (1.69% of positives) or persistent (11.86% of positives) trends less frequently show ZNF671 methylation. Interestingly, patients with HPV 16 infection (52.54% of positives) and >30 years (89.83% of positives) are more likely to appear ZNF671 methylation-positive. However, patients < 30 years with persistent neoplasia (42.86% of positives) also show methylation more frequently. Conclusions: The methylation of ZNF671 is measurable in cervical FFPE material and has prognostic value. Since ZNF671-methylated samples are most likely to be progressing, we recommend the closer monitoring of patients with GynTect^®-positive test results. Full article

This study investigated the morphometric characteristics of adenohypophyseal thyrotrophs and circulating thyroid hormone profiles in dromedary camels (Camelus dromedarius) in relation to age and lactation status. Clinically healthy Brela breed camels were divided into lactating female, and non-lactating female groups across two age categories (5–10 years and ≥11 years), with fifty animals per group. Blood samples were collected before slaughter and pituitary glands were collected post-slaughter and processed for immunohistochemical detection of thyroid-stimulating hormone (TSH) using anti-porcine TSHβ antibody, while morphometric measurements of thyrotrophs were conducted through image analysis. Plasma concentrations of TSH, triiodothyronine (T₃), and thyroxine (T₄) were quantified using validated ELISA and enzyme immunoassay kits. Group differences were analyzed using one-way ANOVA followed by post hoc comparisons, with statistical significance set at p < 0.05. Morphometric analysis revealed that lactating female camels exhibited significantly higher thyrotroph counts compared with non-lactating counterparts, whereas non-lactating females displayed larger cell and nuclear dimensions. Age influenced these patterns, with older camels showing hypertrophied thyrotrophs but reduced functional plasticity compared to younger animals. Plasma hormone assays demonstrated that non-lactating camels had higher TSH and T₄ concentrations, while lactating camels maintained elevated T₃ levels, suggesting enhanced peripheral conversion of T₄ to T₃ during milk production. Additionally, younger camels exhibited higher T₃ concentrations than older animals, indicating age-related decline in thyroidal activity. These findings highlight the dynamic regulation of the hypothalamic–pituitary–thyroid axis in camels, demonstrating how lactation and age shape thyroidal morphology and function to meet diverse physiological demands. These findings not only broaden the comparative endocrinology of underexplored species but also provide physiopathological insights relevant to farm animal management, lactation efficiency, and adaptive metabolism in harsh environments. Full article

Brucellosis, a zoonotic infection caused by the intracellular pathogen Brucella, leads to chronic multi-organ damage. Currently, rapid, accurate, and sensitive diagnostic technologies are crucial for the prevention and control of brucellosis. This study describes the development of a chemiluminescent immunoassay (Bru-CLIA) for sheep and bovine brucellosis antibody detection, utilizing Brucella abortus strain A19 lipopolysaccharide-coated magnetic particles (LPS-MPs) as the serum antigen and acridinium ester-labeled recombinant streptococcal protein G (AE-SPG) for signal generation. After optimizing the assay’s parameters, the Bru-CLIA demonstrated a sensitivity of approximately 1 IU/mL and 2 IU/mL for detecting sheep and bovine brucellosis, respectively. No cross-reactivity was observed with sera from animals immunized with Escherichia coli O157:H7, Mycobacterium tuberculosis, Vibrio cholerae, Legionella, Salmonella, Foot and Mouth Disease virus types O and A, Bovine viral diarrhea virus, Sheep contagious pleuropneumonia, Goat pox virus, or Peste des Petits Ruminants virus, indicating strong specificity. The testing of 81 sheep serum samples and 96 bovine serum samples revealed that Bru-CLIA showed 87.65% and 93.75% concordance with the ID-VET commercial kits for sheep and bovine brucellosis detection, respectively. These results demonstrate that Bru-CLIA offers high specificity, sensitivity, repeatability, and reliability, making it a viable rapid diagnostic tool for the epidemiological surveillance of brucellosis. Full article

Gentamicin (GEN) is a broad-spectrum antibiotic widely used in livestock production, and its excessive residues in animal-derived foods pose potential health risks to consumers. However, conventional colloidal gold immunochromatographic assays (AuNPs-ICA) often suffer from low sensitivity and poor tolerance to sample matrices. Herein, a UiO-66-NH₂ framework decorated with gold nanoparticle (UiO-66-NH₂@Au)-based ICA was employed to construct an ICA platform for GEN detection, combining the optical advantages of AuNPs with the protective and stable octahedral framework of the Metal-organic framework (MOF) to enhance antibody stability under extreme conditions. The method achieved limits of detection for GEN of 0.1 µg/kg in four tested matrices, with recoveries of 80.1–112.0% and coefficients of variation below 11.7%. Compared to traditional AuNPs-ICA, the sensitivity was improved by up to 30-fold, the pH tolerance range was expanded from 6–8 to 4–10, and the organic solvent tolerance to organic solvents expanded up to 40%. Verification with 40 real samples demonstrated excellent correlation (R² > 0.99) with results from commercial ELISA kits. This UiO-66-NH₂@Au-ICA platform offers a new technical solution with high sensitivity, strong good anti-interference performance, and robustness for rapid field detection of GEN residues in products of animal origin and holds significant practical importance for advancing rapid food safety detection technologies. Full article