Search Results (680)

The gamma band observed in human electroencephalography (EEG) has been extensively studied. However, recent research has begun distinguishing the potential roles assigned to phase and non-phase modulation within this band. The primary aim of this study is to analyze the potential role of non-phase gamma modulation in a widely used visual task in human subjects. For this purpose, using a 58-channel EEG recording, gamma activity was evaluated during an oddball task. Responses from 21 healthy subjects were recorded at two separate time points, with an average interval of 49.5 ± 48.9 days. Latency, amplitude, and topographic correlation values were calculated to assess the replicability. Furthermore, potential influence of alpha band harmonics on gamma was analyzed. Topographic analyses revealed a strong negative correlation between gamma phase-locked (synchronous) and non-phase-locked (asynchronous) activity, with correlation coefficients of r < −0.9 for both measures. The results observed between the two time points were robust. The harmonic analysis did not show any potential contribution of the alpha band. The separate analysis of phase and non-phase activity has enabled us to identify distinct roles for each. Establishing non-phase activity as a perceptual “anti-binding” mechanism opens new avenues for exploring a previously unaddressed aspect of gamma activity. Full article

According to the EFSA Report on Zoonoses (2024), yersiniosis was classified as the fourth most commonly reported zoonosis in humans in 2023, with a 13.5% increase in yersiniosis infections compared to 2022. In 2024, the findings were consistent with the 2020–2023 trend. Isolation and identification of enteropathogenic Yersinia is difficult and time consuming, especially when examining food and environmental samples. Among them, Y. pseudoturbeculosis poses a challenge due to the lack of a single selective medium for all bioserotypes. Therefore, faster methods for the detection of Yersinia spp. need to be implemented into the praxis. Rapid identification of pathogens in food or at the time and location of the epidemiological outbreak (point-of-care testing) enables either prevention of the outbreak or early stage diagnosis and prompt decisions. The loop-mediated isothermal amplification (LAMP) is increasingly coming to scientists’ attention as a robust and rapid methodology for pathogen detection in laboratories with limited resources and equipment. The aim of current study is to evaluate, for the first time, the sensitivity of the LAMP protocol based on colorimetric detection in the visible spectrum in comparison with that of the digital droplet PCR (ddPCR). For this aim, a series of decimal logarithmic dilutions of the pathogen Y. pseudotuberculosis in artificially contaminated raw goat milk was used. One commercial LAMP kit with two different dyes (one dsDNA-binding and one Mg²⁺-sensitive) was compared to the sensitivity of the detection to ddPCR. The results obtained revealed a high sensitivity of the kit for detection of DNA isolated from artificially contaminated milk samples in the following range: visible detection based on visible color change—3.1 × 10⁴ mL (violet dye) and 3.4 × 10³/mL (blue dye); detection with gel electrophoresis—2.0 × 10¹/mL (violet dye) and 3.4 × 10²/mL (blue dye). The enumeration of the DNA copies in the same samples was performed with ddPCR, with a detection limit of 2.0 × 10¹/mL. Our results indicate the potential and the possible applicability of the LAMP method for rapid and sensitive visual detection of Y. pseudotuberculosis in raw goat milk. The presented ddPCR protocol can be used for highly sensitive identification and enumeration of Y. pseudtuberculosis in raw goat milk. In conclusion, the conducted comparison is of importance for future implementation of LAMP protocols for on-field analysis near the sampling site and point-of-care or laboratory diagnostics of Y. pseudtuberculosis after the successful validation procedure of an appropriate LAMP protocol. Full article

Photodynamic therapy (PDT) is a technique based on the use of photosensitizers activated by light to destroy cancer cells in the presence of oxygen. This enables localized cancer treatment and, in some settings, fluorescence-guided visualization. However, the efficacy and clinical translation of PDT have been limited by the low specificity of traditional photosensitizers. The aim of the study is to create a ligand-guided PDT approach for pancreatic ductal adenocarcinoma (PDAC) using a peptide-conjugated photosensitizer binding to integrin αvβ6, which is a receptor linked to tumor growth and prevalent in PDAC cells. Current treatment options for this tumor are limited, with surgical resection and chemotherapy only effective when the tumor is detected early. Given the limited treatment options for PDAC, PDT via αvβ6 offers a new pathway for precision treatment. The cyclic peptide cyclo[FRGDLAFp(NMe)K], recognized for its high affinity to αvβ6, was chosen to guide a phthalocyanine-class photosensitizer toward αvβ6-expressing PDAC models. The PDT approach was further refined by developing 3D spheroid models and in vivo BxPc3 xenograft models in NOD/SCID mice, where its therapeutic efficacy was assessed. In the absence of a non-targeted control photosensitizer, a contribution from non-specific accumulation and EPR effects in the in vivo setting cannot be fully ruled out. This study highlights the potential of a peptide-guided photosensitizer, demonstrating uptake and photodynamic activity in spheroids, with moderate in vivo results addressing tumor microenvironment challenges. Optimization of PDT dosing, laser precision, and preclinical models, such as patient-derived xenografts, are crucial to enhance clinical translation. Full article

B7-H3 (CD276), an immune checkpoint protein, is overexpressed in malignant tumors, while its expression in normal tissues is low, and several B7-H3-targeting therapies are under clinical evaluation. Radionuclide molecular imaging offers a non-invasive method for determining B7-H3 expression levels and may aid in improved patient selection. The feasibility of the use of Affibody molecules for the visualization of B7-H3 was demonstrated earlier. The selection of an approach for routine labeling providing high radiochemical yields and reproducibility is, however, critical for successful clinical translation. The optimal combination of a targeting protein, chelator/linker, and radionuclide should provide high-contrast visualization. In this study, we evaluated an acyclic chelator, tris(3,4-hydroxypyridinone) (THP), for labeling of the Affibody molecule Z_{B7-H3_2} with ⁶⁸Ga and compared its impact on radiolabeling efficiency and targeting properties with the impact of the cyclic chelator 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA). Maleimide derivatives of THP and NOTA were site-specifically coupled to the C-terminal cysteine of Z_{B7-H3_2}. THP-Z_{B7-H3_2} was successfully labeled with ⁶⁸Ga within 5 min of incubation at room temperature, achieving a 100% radiochemical yield. NOTA-Z_B7-H3 required heating at 60 °C for 10 min, and the radiochemical yield was lower. Both radioconjugates exhibited specific binding to B7-H3-expressing cells with similar binding strength, and both tracers demonstrated similar tumor uptake (p > 0.05) in a murine model. The biodistribution was similar, although [⁶⁸Ga]Ga-NOTA-Z_{B7-H3_2} provided slightly but significantly higher tumor-to-liver and tumor-to-spleen ratios. Nonetheless, the advantages of THP include rapid and mild radiolabeling with high efficiency, eliminating the need for heating and a post-purification step, which suggests a potential for streamlined clinical translation of Z_{B7-H3_2}. Full article

ALS is a severe neuromuscular disease classically characterized by the progressive loss of motor neurons, leading to incremental muscle weakness and eventually death. Current treatment options for ALS have proven to have limited effect, merely delaying the progression of symptoms and prolonging patient survival. This motor neuron subtype-related differential vulnerability has been linked to neuron excitability, metabolism, and protein aggregation. Calcium dysregulation, which serves as an important second messenger in neural signaling pathways, has been implicated in each of these mechanisms and represents a potential target for therapeutic intervention. Armed with cutting-edge tools for visualizing and recording calcium transients in vivo, ALS researchers have delved deeper into the role of calcium dysregulation in disease in recent years. Vulnerable motor neuron populations display an excess of calcium-permeable ion channels together with reduced expression of calcium-binding proteins, generating a cellular environment primed for excitotoxic stress. Loss of inhibitory synaptic input further heightens susceptibility to calcium overload. Paradoxically, some evidence suggests that elevated neuronal activity can exert neuroprotective effects, highlighting the complexity of activity-dependent calcium signaling in ALS. Additionally, ALS-related toxic protein accumulation disrupts calcium homeostasis, contributing to endoplasmic reticulum stress and mitochondrial dysfunction. Emerging data indicate that calcium dysregulation impairs neuron-glia communication, amplifying neuroinflammation and accelerating disease progression. This review aims to synthesize current evidence on how calcium imbalance contributes to motor neuron vulnerability and degeneration in ALS. By exploring the cellular, synaptic, and network-level mechanisms of calcium dysregulation in ALS, the review examines its interplay with mitochondrial and ER stress and explores its impact on neuron-glia interactions with the aim of synthesizing key mechanistic insights into the disease pathogenesis and therapeutic targets. Full article

Background: Radiolabeled fibroblast activation protein inhibitors (FAPIs) have emerged as novel radiopharmaceutical agents for tumor diagnosis. Compared with [¹⁸F]fluoro-2-deoxy-D-glucose ([¹⁸F]FDG), which reflects glucose uptake in metabolically active regions, FAPIs mainly bind to the fibroblast activation protein (FAP), which is highly expressed in tumor-associated fibroblasts, forming a pronounced signal. Several studies suggested potential superiority of FAPI tracers above [¹⁸F]FDG-based imaging in a variety of tumor entities. In this systematic review, we focus on the comparison of FAPI-PET/CT and [¹⁸F]FDG-PET/CT in upper-abdominal tumors. Methods: Original research published from 1 January 2021 to 22 December 2024 was collected from the PubMed and Web of Science databases (CRD42025648267). This research included only clinical studies, excluding conference abstracts and case reports. The risk of bias was assessed with the QUADAS-2 tool, and all evaluation steps performed independently by three independent reviewers. A systematic quality assessment of the included studies was conducted based on the imaging performance of FAPI-PET/CT and [¹⁸F]FDG-PET/CT for pancreatic, liver, and gastric cancers. The meta-analysis used relative risk (RR) as the effect size, with bias assessed via the Peters test (p-value > 0.05). Cochran’s Q test and I-squared value are used to comprehensively evaluate the magnitude of heterogeneity. Analyses and data visualization were performed in R language. Results: The database search identified 3272 articles. After screening, 31 studies were included in this analysis. The original studies enrolled 1377 participants (M/F: 850/527; ages predominantly between 50 and 70). Of these, 939 patients were ultimately diagnosed with tumors (five cancer subtypes) and included in this analysis. Meta-analysis results showed that FAPI-PET/CT significantly surpassed [¹⁸F]FDG-PET/CT in the detection of primary lesions (RRs = 1.20 and 1.17), lymph nodes (RRs = 1.18 and 1.24), distant metastases (RRs = 1.22 and 1.51), peritoneal metastases (RRs = 1.31 and 2.22), and bone metastases (RRs = 1.16 and 1.23). The two imaging methods exhibit clear differences in diagnostic performance (sensitivity: 98% vs. 79%; specificity: 83% vs. 87%), and FAPI-PET/CT demonstrates high and stable diagnostic performance (RRs = 1.20 and 1.17). Conclusions: Compared with [¹⁸F]FDG-PET/CT, FAPI-PET/CT demonstrates significant advantages in detecting primary lesions, lymph nodes, distant metastases, and peritoneal and bone metastases in pancreatic, liver, and gastric cancers (RR > 1.0). Overall, FAPI-PET/CT shows better diagnostic performance (RR > 1.0). Full article

Sulfamethazine (SMZ) is widely used in livestock production, and its residues can enter water and soil environments, posing potential risks to human health and ecosystems. This study focuses on environmental samples and constructs an AuNP-based colorimetric aptasensor using the SMZ1S aptamer for the rapid visual detection of SMZ. Under optimized conditions, the aptasensor showed a wide linear range from 0.05 to 0.4 µg/mL and a limit of detection of 0.039 µg/mL. Molecular dynamics simulations have demonstrated that the aptamer’s binding to SMZ is stable, providing a theoretical basis for the high selectivity of the aptasensor. Spike-and-recovery experiments yielded recoveries of 87.3–105.5%, 88.6–102.8%, and 87.5–103.4% for SMZ in lake water, tap water, and soil samples, respectively, with relative standard deviations of 5.9–8.3%, 8.0–10.6%, and 4.8–9.6%, showing good agreement with high-performance liquid chromatography (HPLC) results (R² ≥ 0.981). Overall, the proposed aptasensor provides a simple and effective approach for rapid detection of SMZ in environmental samples. Full article

In on-site rapid detection, the electrochemical method boasts high sensitivity and rapid response capabilities, while the colorimetric method can provide intuitive visual readings suitable for on-site screening. Therefore, this study developed an innovative dual-mode electrochemical/colorimetric aptasensor for the accurate detection of malathion (MAL) in vegetables. The sensor combines magnetic Fe₃O₄@ZIF-8-DNA composites and CuZr-MOF-cDNA probes, enabling simultaneous detection of the target through electrochemical reactions and colorimetric changes. The introduction of CuZr-MOF not only enhances the sensor’s conductivity but also significantly amplifies the electrochemical signal through its catalytic properties. The magnetic Fe₃O₄@ZIF-8-DNA composite facilitates solid–liquid separation under an external magnetic field. When the target MAL is present, the aptamer binds to the target, causing the CuZr-MOF-cDNA probes to release from the composite, altering the number of free probes in the supernatant and generating varying intensities of colorimetric signals. Meanwhile, the MAL captured in the precipitate by the aptamer is quantitatively detected through electrochemical methods. Experimental results demonstrate that as the target concentration increases, the colorimetric signal intensifies while the electrochemical signal weakens, showing a good linear relationship between the two. The aptasensor’s limit of detection (LOD) for colorimetric and electrochemical modes was 1.57 × 10⁻¹¹ M and 4.76 × 10⁻¹¹ M, respectively, with recoveries ranging from 87.71% to 107.68% and relative standard deviations between 3.23% and 10.75%. This method exhibits high sensitivity, excellent selectivity, and strong reliability, providing a novel technique for the accurate quantification of MAL in vegetables, particularly suited for on-site rapid detection. Full article

Thiazides are among the most efficacious and commonly used drugs for the treatment of hypertension. The nanomolar stabilizer N3SA binds specifically to the recently discovered thiazide-binding site of the membrane target NAPE-PLD, showing sustained arterial blood pressure-lowering effects and vasodilation in spontaneous hypertensive rats (SHRs). To further support the relation between stabilizers anchored to NAPE-PLD and their beneficial effects on hypertension, we selected compound analogues of N3SA with chemical modifications at the three target-interacting sulfonic groups, including the drug Suramin. Each compound was injected i.v in an adult SHR (systolic blood pressure of 217 ± 5 mmHg) to evaluate the frequency components contribution to cerebral and peripheral arteriolar vasomotion. We visualized the pial and rectus femoral muscle microcirculation by Epi-illumination, measuring changes in the rhythmic arteriolar diameter. Findings showed that the minor structural differences in compounds correlated with the contribution of the six different frequency components affecting the arterial tone, as well as their vasodilatory effects, in both cerebral and femoral muscle arterioles. These results provide evidence that the spectra analysis of the regulation mechanisms of vascular tone and arterial blood pressure can accurately reflect the structure–activity correlations of different analogues of an antihypertensive compound. Full article

Background/Objectives: Colorectal cancer (CRC) is a leading cause of cancer morbidity and mortality worldwide. Despite therapeutic advances, prevention through dietary bioactives remains a promising strategy. The Annurca apple (Malus pumila Miller cv. Annurca), a Mediterranean food rich in chlorogenic acid, exhibits antioxidant and anti-inflammatory effects. This study evaluated, via molecular docking, the multi-target interaction profile of chlorogenic acid against key CRC-related proteins. Methods: The optimized 3D structure of chlorogenic acid was docked to ten protein targets implicated in CRC pathogenesis, using the GOLD v.2022.3.0 software. Validation of the docking protocol was achieved by re-docking native ligands (RMSD ≤ 2.0 Å). Binding affinities were assessed by ChemPLP scoring, and interaction networks were visualized in Maestro Schrödinger. Results: Chlorogenic acid displayed consistent binding across all evaluated targets (ChemPLP 57.12–69.66), showing the highest affinity for nAChR (69.66), CXCR2 (65.13), ERβ (63.18) and TGFBR2 (62.94). The ligand formed multiple hydrogen bonds and π-π stacking interactions involving Asp1040 (VEGFR-1), Cys919 (VEGFR-2), Lys320 (CXCR2), and Tyr195 residues (nAChR), contributing to strong complex stabilization. Interaction patterns in CYP19A1, ERβ, and ERRγ suggested potential modulation of hormonal and metabolic signaling. The compound also demonstrated stable binding to mTOR (60.01), indicating a possible inhibitory role in proliferative pathways. Collectively, these findings reveal a broad, polypharmacological binding profile involving angiogenic, inflammatory, and hormonal regulators. Conclusions: Chlorogenic acid acts as a promising multi-target ligand in CRC prevention, with our in silico evidence supporting its ability to modulate diverse oncogenic pathways. Further experimental studies are warranted to confirm its efficacy and translational potential. Full article

Surface-imprinted polymer (SIP)-based biomimetic sensors are promising for direct whole-bacteria detection; however, the commonly used fabrication approach (micro-contact imprinting) often suffers from limited imprint density, heterogeneous template distribution, and poor reproducibility. Here, we introduce a photolithography-defined master stamp featuring E. coli mimics, enabling high-density, well-oriented cavity arrays (3 × 10⁷ imprints/cm²). Crucially, the cavity arrangement is engineered such that the SIP layer functions simultaneously as the bioreceptor and as a diffraction grating, enabling label-free optical quantification by reflectance changes without additional transduction layers. Finite-difference time-domain (FDTD) simulations are used to model and visualize the optical response upon bacterial binding. Proof-of-concept experiments using a differential two-well configuration confirm concentration-dependent detection of E. coli in PBS, demonstrating a sensitive, low-cost, and scalable sensing concept that can be readily extended to other bacterial targets by redesigning the photolithographic master. Full article

Bentonite is a multifunctional material widely used in industry, agriculture, food processing, and medicine due to its strong binding and absorption properties. This study investigates the effects of bentonite pre-treatment at different concentrations (0.5–5%) on soybean sprout growth and nutritional quality. Moderate levels, particularly 1–3% (BP-1 and BP-3), significantly increased sprout yield (up to 16.1%) and vitamin C content (up to 18.91 mg/100 g FW), while maintaining desirable moisture and visual quality. Color evaluation showed higher yellowness (b*), suggesting improved consumer appeal. Mineral profiling indicated substantial enhancement of essential minerals across treatments, with the highest total mineral content recorded in the BP-5 group. Phosphorus, potassium, copper, and iron were notably enriched; however, elevated copper and reduced zinc at higher concentrations indicate potential nutritional risk. Isoflavone analysis revealed increased total isoflavones, especially glucoside forms such as daidzin and genistin, while aglycones like genistein decreased, reflecting bentonite’s selective influence on isoflavone metabolism. Antioxidant properties—including DPPH scavenging capacity, total polyphenols, flavonoids, and SOD-like activity—were significantly enhanced. Amino acid profiling also showed increases in essential amino acids, including methionine and valine, along with higher γ-aminobutyric acid (GABA). Overall, bentonite demonstrates strong potential as a natural additive for improving soybean sprout productivity and functional quality, with the most favorable outcomes observed at 1–3% concentrations. Full article

The bHLH transcription factors play a crucial regulatory role in plant growth and development. In this study, the CcbHLH68 gene was cloned from the pepper cultivar ‘Shuanla’. Subsequent bioinformatics analysis, subcellular localization, expression pattern profiling, along with yeast one-hybrid and dual-luciferase reporter assays, were conducted to preliminarily elucidate its regulatory mechanism in capsaicinoid biosynthesis. The results revealed that the visualization of upstream cis-elements of CcbHLH68 suggests its potential regulation by hormones. Furthermore, subcellular localization experiments confirmed that the CcbHLH68 protein is localized in the nucleus. Expression analysis of CcbHLH68 across different tissues by qRT-PCR identified its predominant expression in the placenta at 30 days post-anthesis. Further experimental evidence from both gene silencing and transient overexpression assays demonstrated a positive correlation between CcbHLH68 and the expression of multiple capsaicinoid biosynthetic genes. When it was silenced or transiently overexpressed, the content of capsaicinoids decreased by 40.9% or increased by 113.7%, respectively. Yeast one-hybrid and dual-luciferase reporter assays confirmed that CcbHLH68 can directly bind to the CcCOMT promoter and activate its transcription. In summary, this study preliminarily reveals the molecular mechanism by which CcbHLH68 participates in capsaicinoid biosynthesis through regulating the expression of key genes in the biosynthetic pathway, thereby providing a theoretical foundation for enhancing capsaicinoid content via molecular breeding. Full article

P-glycoprotein (P-gp) is an ATP-driven transporter that effluxes a wide range of xenobiotics from cells, and its overexpression is a primary cause of multidrug resistance (MDR) in cancer. It is well-established that P-gp functions through conformational changes, yet its large-scale structural dynamics at work have been unexplored. Here, we directly visualized single P-gp molecules reconstituted in nanodiscs using high-speed atomic force microscopy (HS-AFM). The HS-AFM movies revealed that P-gp is intrinsically dynamic in its apo state, with its nucleotide-binding domains (NBDs) undergoing large, spontaneous opening and closing motions. However, addition of ATP stabilized a conformation characterized by NBD proximity with a strong tendency toward closure. We then leveraged this dynamic viewpoint to elucidate the relationship between Elacridar’s function and the resulting structural dynamics of P-gp. Elacridar is designed to overcome multidrug resistance (MDR) in cancer and acts as a potent dual inhibitor of both P-gp and the Breast Cancer Resistance Protein (BCRP), effectively blocking the drug efflux function of these transporters. This inhibitor has suggested concentration-dependent function: it is effluxed as a substrate at low concentrations and acts as an inhibitor at high concentrations. Our direct observations revealed that low concentrations induced active dynamics in P-gp, whereas high concentrations severely restricted its motion, leading to a rigid, non-productive state. Our study provides critical insights into how observing molecular motion itself can unravel complex biological mechanisms. Full article

This study aimed to identify novel angiotensin-converting enzyme (ACE)-inhibitory peptides from royal jelly proteins (RJPs) by integrating in silico digestion, virtual screening, and in vitro evaluation. Three major royal jelly proteins (MRJP1-3) were subjected to in silico digestion using 16 enzymatic systems, yielding 1411 unique peptides. Virtual screening based on predicted bioactivity, toxicity, water solubility, and ADMET profiles resulted in the selection of 27 candidate peptides. Molecular docking revealed strong binding affinities for these peptides compared with the positive control captopril, among which PYPDWSFAK and RPYPDWSF exhibited potent ACE-inhibitory activity, with IC₅₀ values of 110 ± 1.02 μmol/L and 204 ± 0.61 μmol/L, respectively. Kinetic analysis indicated that PYPDWSFAK acts as a mixed-type ACE inhibitor. Docking visualization demonstrated that PYPDWSFAK forms multiple hydrogen bonds with key residues in the ACE active pocket and directly coordinates with the catalytic Zn²⁺ ion. Cellular assays showed that PYPDWSFAK was non-cytotoxic, suppressed Ang II–induced endothelial cell migration, restored NO and ET-1 balance, and enhanced SOD and GSH-Px activities. Overall, this study enriches the repertoire of ACE-inhibitory peptides derived from royal jelly proteins. Furthermore, PYPDWSFAK is identified as a promising ACE-inhibitory peptide with potential for incorporation into natural antihypertensive ingredients or functional foods. Full article