Search Results (1,802)

Objectives: Anterior petrosectomy is a challenging neurosurgical procedure requiring precise identification and preservation of multiple critical structures. This technical note explores the feasibility of using real-time near-infrared indocyanine green (NIR-ICG) fluorescence with white light overlay to enhance visualization of the petrous internal carotid artery (ICA) during transpetrosal drilling. We aimed to assess its utility for planning and performing modified Dolenc–Kawase drilling. Methods: We integrated NIR-ICG and white light overlay using a robotic microscope with simultaneous visualization capabilities. This technique was applied to improve neurovascular preservation and skull base landmark identification. Intraoperative video frames and images were captured during an anterior transpetrosal approach for a petroclival meningioma, with technical details, surgical time, and feedback documented. Results: Real-time NIR-ICG with white light overlay successfully identified the posterior genu, horizontal petrosal segment, anterior genu, and superior petrosal sinus. It facilitated precise localization of cochlear landmarks, enabling tailored drilling of the Dolenc–Kawase rhomboid according to patient anatomy and accommodating potential anatomical variants. Conclusions: This approach could enhance intraoperative safety and improve exposure, possibly reducing neurovascular risks without extending operative time. It may serve as a valuable adjunct for complex skull base surgeries. Full article

Harmful algal blooms (HABs) caused by toxic species such as Pseudo-nitzschia hasleana pose significant risks to marine ecosystems and human health. This study investigates the effects of heating rate on the fluorescence temperature curves (FTCs) of P. hasleana and compares them with non-toxic species (Phaeodactylum tricornutum and Picochlorum maculatum) to design a reliable detection method. An increasing heating rate leads to a change in the temperature spectrum of the fluorescence of the studied algae and to increasing differences between them. During the study, the FTCs were measured in the temperature range of 20–80 °C and at heating rates of 1, 2, 3, and 6°/min. The results showed that P. hasleana exhibited a distinct local fluorescence maximum at 45–55 °C when heated at a rate of 3 °C/min or more, which was absent in non-toxic species. Additionally, rapid heating (6 °C/min) preserved fluorescent pigment–protein complexes, yielding four-fold higher fluorescence intensity at 70–80 °C compared to slower rates. There were no such changes for the microalgae P. maculatum and P. tricornutum. The results of this study make it possible to increase the efficiency of detecting hazardous microalgae using non-invasive optical monitoring methods. These findings demonstrate that controlled heating protocols can enhance the species-specific identification of toxic microalgae, offering a practical tool for early HAB detection. Full article

Amphorae are a significant type of Roman pottery, serving as both transport containers and indicators of economic and technological practices across the Empire. Despite their importance in interpreting local economic dynamics in Roman Lusitania, the production origins, technological characteristics and functional roles of amphorae from Conímbriga in Portugal remain poorly understood. Previous research has focused primarily on typological comparisons with imported forms, providing limited insight into whether these vessels were produced locally or how they were incorporated into regional trade and agricultural systems. This study takes a multi-analytical approach to examine a group of ten amphorae dating from the 1st to the 5th century AD. While these vessels formally resemble well-known types from Gaul and Hispania, they appear to have been manufactured locally using fabrics consistent with those found in regional common wares. To examine the raw materials, production techniques and possible contents, the analysis integrates a typological assessment with a range of archaeometric methods, including optical microscopy (OM), X-ray diffraction (XRD), X-ray fluorescence (XRF) and scanning electron microscopy with energy-dispersive spectroscopy (SEM-EDS). Recent gas chromatography–mass spectrometry (GC–MS) results contextualise these data further, enabling an exploration of how specific technological choices may have influenced or reflected the intended function of the amphorae. The findings shed new light on local ceramic production and contributed to broader debates concerning Roman provincial economies and material culture. Full article

The accumulation of high levels of manganese ions complexed with small molecules has been proposed as a pivotal factor contributing to the extraordinary radiation resistance of Deinococcus radiodurans. However, the molecular mechanisms governing the manganese ion homeostasis remain elusive. In this study, we characterize the role of the MntABC transporter system for Mn ion accumulation in D. radiodurans. Its cellular membrane localization is unequivocally demonstrated through fluorescence labeling techniques. Mutation of the protein components of the MntABC led to a significant decrease in intracellular Mn ion accumulation, concomitant with impaired cellular growth, decreased resistance against hydrogen peroxide, and gamma-ray irradiation-induced oxidative stresses, indicating that the MntABC system plays an indispensable role in resistance of D. radiodurans to oxidative stresses. Protein structure modeling and molecular docking are employed to analyze the key active sites of the MntABC proteins and their intermolecular interactions. The results demonstrate that the MntABC system is essential for maintaining Mn ion homeostasis and the oxidative stress resistance of D. radiodurans. Full article

Sodalite is a common feldspathoid in alkaline systems, with some varieties exhibiting notable fluorescence due to impurity activators. This study reports the first documented occurrence and characterization of fluorescent sodalite from the classic Ditrău Alkaline Massif, Romania, where its optical properties were previously undescribed. Sodalite-bearing syenite samples from different perimeters of the massif were investigated using macroscopic UV fluorescence, petrographic microscopy, and vibrational spectroscopy (Raman and FT-IR). The sodalite occurs as a late-stage, interstitial and poikilitic mineral, often associated with alteration to cancrinite. Under long-wave UV (365 nm) light, it exhibits spatially variable fluorescence, from absent in parts of the western Prişca perimeter to strong, uniform orange in the eastern Aurora perimeter. Raman and FT-IR spectroscopy confirmed the mineral’s identity and revealed subtle spectral variations, particularly the presence of a minor cancrinite component in some analyses. The vibrant orange fluorescence is consistent with activation by disulfide radical anion ($S_2^{·-}$) activators, formed in the sulfur- and chlorine-rich late-stage fluids characteristic of the massif’s evolution. The geographic variation in fluorescence intensity serves as a potential indicator of the geochemical heterogeneity of these fluids across the massif, linking the strongest fluorescence to the most evolved portions of the igneous complex. This finding opens a new avenue for using fluorescence as a tool for petrogenetic investigation in this classic locality. Full article

Insights into the state of individual cells within a living organism are essential for identifying diseases and abnormalities. The internal state of a cell is reflected in its morphological features and changes in the localization of intracellular molecules. Using this information, it is possible to infer the state of the cells with high precision. In recent years, technological advancements and improvements in instrument specifications have made large-scale analyses, such as single-cell analysis, more widely accessible. Among these technologies, imaging flow cytometry (IFC) is a high-throughput imaging platform that can simultaneously acquire information from flow cytometry (FCM) and cellular images. While conventional FCM can only obtain fluorescence intensity information corresponding to each detector, IFC can acquire multidimensional information, including cellular morphology and the spatial arrangement of proteins, nucleic acids, and organelles for each imaging channel. This enables the discrimination of cell types and states based on the localization of proteins and organelles, which is difficult to assess accurately using conventional FCM. Because IFC can acquire a large number of single-cell morphological images in a short time, it is well suited for automated classification using machine learning. Furthermore, commercial instruments that combine integrated imaging and cell sorting capabilities have recently become available, enabling the sorting of cells based on their image information. In this review, we specifically highlight practical applications of IFC in four representative areas: cell cycle analysis, protein localization analysis, immunological synapse formation, and the detection of leukemic cells. In addition, particular emphasis is placed on applications that directly contribute to elucidating molecular mechanisms, thereby distinguishing this review from previous general overviews of IFC. IFC enables the estimation of cell cycle phases from large numbers of acquired cellular images using machine learning, thereby allowing more precise cell cycle analysis. Moreover, IFC has been applied to investigate intracellular survival and differentiation signals triggered by external stimuli, to monitor DNA damage responses such as γH2AX foci formation, and more recently, to detect immune synapse formation among interacting cells within large populations and to analyze these interactions at the molecular level. In hematological malignancies, IFC combined with fluorescence in situ hybridization (FISH) enables high-throughput detection of chromosomal abnormalities, such as BCR-ABL1 translocations. These advances demonstrate that IFC provides not only morphological and functional insights but also clinically relevant genomic information at the single-cell level. By summarizing these unique applications, this review aims to complement existing publications and provide researchers with practical insights into how IFC can be implemented in both basic and translational research. Full article

The primary mechanism of non-steroidal anti-inflammatory drugs (NSAIDs) is inhibition of prostaglandin production mediated by cyclooxygenase. Given the possible association of cyclooxygenase-2, but not cyclooxygenase-1, with membrane lipid rafts, we assessed whether the lipid raft membrane interactivity of NSAIDs correlates with cyclooxygenase-2 selectivity. Lipid raft model membranes and reference membranes were prepared with 1,2-dioleoylphosphatidylcholine/sphingomyelin/cholesterol and 1,2-dipalmitoylphosphatidylcholine, respectively. After treating the membranes with 2–50 μM NSAIDs at pH 7.4, 6.5, and 5.5, fluorescence polarization was measured to determine their membrane interactivity. Conventional NSAIDs (diclofenac, ibuprofen, indomethacin, aspirin, and flurbiprofen) and Coxibs (lumiracoxib, etoricoxib, celecoxib, valdecoxib, and rofecoxib) decreased membrane fluidity, whereas Oxicams (meloxicam, piroxicam, tenoxicam, and lornoxicam) increased. Membrane effects of NSAIDs were so dependent on medium pH that they significantly increased with reducing pH from 7.4 to 5.5. Under inflammatory acidic conditions, the lipid raft membrane interactivity of NSAIDs was more likely to correlate with cyclooxygenase-2 selectivity than the reference membrane interactivity. It is hypothesized that NSAIDs may interact with lipid raft membranes to induce membrane fluidity changes with the potency corresponding to cyclooxygenase-2 inhibition, disrupting the structural and functional integrity of lipid rafts to affect the activity of cyclooxygenase-2 localized in lipid rafts, resulting in cyclooxygenase-2 selective inhibition. Full article

Addressing the critical challenge in the differential diagnosis of severe inflammatory lung diseases, we propose a novel methodology for the analysis of macrophage surface receptors, CD68 and CD206, using specific non-antibody ligands. We developed a non-antibody alternative for the fluorometric detection of CD68+ cells, focusing on macrophages as key functional markers in inflammatory processes. Our marker based on dioleylphosphatidylserine (DOPS), a specific ligand to CD68, was incorporated into a liposomal delivery system. The specificity of this DOPS-based ligand can be precisely modulated by the liposome’s composition and the polyvalent presentation of the ligand. We synthesized a series of fluorescently-labeled DOPS-based ligands and developed a liposome-based sandwich fluorometric assay. This assay enables the isolation and quantification of CD68 receptor presence from bronchoalveolar lavage fluid (BALF). The results confirmed the specific binding of DOPS/lecithin liposomes to CD68+ cells compared to control lecithin systems. Furthermore, the incorporation of PEGylated ‘stealth’ liposomes significantly enhanced binding specificity and facilitated the generation of distinct binding profiles, which proved valuable in differentiating various inflammatory conditions. This approach yielded unique binding profiles of PS-based ligands to CD68+ cells, which varied significantly among a broad range of respiratory conditions, including primary ciliary dyskinesia, bronchial asthma, bronchitis, bacterial infection, pneumonia, and bronchiectasis. Confocal Laser Scanning Microscopy demonstrated selective binding and intracellular localization of the DOPS-based marker within CD68+ macrophages from BALF samples of patients with bronchitis or asthma. The binding parameters of this multivalent composite ligand with the CD68 receptor are comparable to those of antibodies. The inherent binding specificity of phosphatidylserine may offer a sufficient and viable alternative to conventional antibodies. Our results demonstrate the remarkable potential of this novel DOPS-based assay as a complementary tool for the developing non-antibody-based systems for the differential diagnosis of the respiratory diseases, warranting further investigation in larger clinical studies. Full article

Indocyanine green (ICG) is a fluorescent agent which is characterized by a wide range of applications in the proper visualization of the operating field, differentiation of vital structures, and localization of lesions to be excised. An investigation and overview of novel approaches of indocyanine green in modern gynecological oncology was conducted, including ovarian cancer surgery with its compartmental approach and compartmental surgery in endometrial cancer. Ureteral visualization and perfusion, lymphography, lymph node transfers, or the localization of anastomotic leakage in bowel surgery are examples of applications aimed at reducing the risk of surgical complications and improving the patients’ quality of life. The general use of indocyanine green in lymph node detection, subcategorized and analyzed, is constantly improved and reviewed. A therapeutic approach with macromolecules is being tested in preclinical models. Early results could suggest the future application of indocyanine green not only in broad-sense imaging but also as a cytotoxic agent conjugated with macromolecules. Further studies on the application of indocyanine green in laparoscopy, open surgery, and finally as a curative cytotoxic agent are needed. Full article

Dithiocarbamate fungicides, including benomyl (methyl 1-butylcarbamoyl-2-benzimidazolecarbamate), share a common mechanism of toxicity by inhibiting aldehyde dehydrogenases (ALDHs), enzymes essential for detoxifying reactive aldehydes. One such aldehyde, 3,4-dihydroxyphenylacetaldehyde (DOPAL), a dopamine metabolite, is implicated in the catecholaldehyde hypothesis of Parkinson’s disease. This study examines ALDH inhibition as the molecular initiating event (MIE) within an adverse outcome pathway (AOP) leading to neurotoxicity. Caenorhabditis elegans at the L4 stage were exposed for 24 h to 10 or 100 μM benomyl. While 10 μM had no significant effect on lethality, growth, or reproduction, 100 μM induced adverse effects, albeit with low lethality. Both doses inhibited ALH activity, an effect mitigated by Alda-1, a selective ALDH activator. Alda-1 alone increased ALH-1 protein levels but did not alter benomyl-induced protein localization and relative abundance. Benomyl exposure also elevated oxidative stress markers—superoxide dismutase, catalase, and lipid peroxidation—which Alda-1 reduced. Neurotoxicity was evidenced by dopaminergic dysfunction, including impaired basal slowing response, neuronal morphological abnormalities, and reduced locomotion upon optogenetic activation. Fluorescent reporter assays confirmed ALH-1 presence in dopaminergic neurons. These results identify ALH inhibition as the MIE in benomyl-induced neurotoxicity, linking dopaminergic degeneration and redox imbalance to the catecholaldehyde hypothesis, and providing mechanistic insights into an AOP relevant to neurodegenerative disorders. Full article

Upon activation, neutrophils perform three distinct functions: phagocytosis, degranulation of antimicrobial substances into the extracellular medium, and release of neutrophil extracellular traps. Determination of the nuclear area expansion of neutrophils activated to release neutrophil extracellular traps has become critical in demonstrating early neutrophil activation and has become standard. Here, we demonstrate an automated method for measuring nuclear area expansion in two different mammalian species: canine and bovine. For both species, neutrophils were isolated from peripheral blood and co-incubated with fresh spermatozoa for up to 120 min for canine neutrophil–spermatozoa and recently thawed cryopreserved spermatozoa up to 240 min for bovine neutrophil–spermatozoa. Fluorescence images were acquired using a TissueFAXS microscope and then analyzed using StrataQuest v.7.0 software. The images show the release of neutrophil extracellular traps upon activation with spermatozoa for both species, as evidenced by the co-localization of neutrophil elastase and DNA staining. Neutrophil nuclei were expanded as early as 15 min and were detected at up to 120 min in both species. Analysis by nuclei segmentation showed that the data sets generated for both species were reliable and consistent with previously published methods. The method was developed as an automated alternative for measuring the area expansion of neutrophil nuclei in different species. Full article

This paper presents the results of a comprehensive study of the wear processes of press tools used in the molding of uranium dioxide (UO₂) nuclear fuel pellets. Particular attention is paid to the analysis of the influence of operating conditions on changes in microstructure, geometry and physical and mechanical properties of working surfaces of molds and punches. The studies using scanning electron microscopy (SEM), X-ray fluorescence (XRF) and X-ray phase analysis (XRD) methods, as well as evaluation of microhardness and roughness, allowed to identify the dominant failure mechanisms—abrasive and adhesive wear, microcrack formation and local degradation of coatings. The results of the experiments confirmed the presence of progressive changes on the working surfaces of the tool, affecting the formation of defects of fuel pellets and reducing the service life of the press equipment. This work allows us to not only better understand the wear patterns in the batch production of nuclear fuel, but also to formulate practical recommendations to increase tool life by optimizing pressing modes and using wear-resistant coatings. Full article

Cohesin organizes the genome into spatially segregated loops and topologically associated domains by loop extrusion. In addition, it ensures cohesion of sister chromatids after replication. Thus, cohesin is expected to limit chromatin dynamics by ensuring cohesion and compacting chromatin in the interphase. Nonetheless, loop extrusion is an example of chromatin dynamics; thus, cohesin could promote the dynamics of genomic loci at the scale of individual loops and contact domains. Moreover, given that the extruding activity of cohesin after replication is supplemented by its cohesive activity, the impact of cohesin on chromatin dynamics in different phases of the cell cycle may vary. Of particular interest is the cohesin’s role in the regulation of the dynamics of damaged chromatin, which remains insufficiently studied. Here, we visualized a genomic locus using the CRISPR-Sirius system in human cells with auxin-induced depletion of the cohesin subunit RAD21. Cohesin depletion increased the local spatial dynamics of the visualized locus on a time scale of fractions of a second to one minute. This effect was observed in both replicated and unreplicated chromatin. However, the increase in the mobility of the visualized locus upon cohesin depletion was more pronounced in the former. In addition, we showed that cohesin depletion did not affect the local mobility of double-strand break repair foci visualized using a fluorescent fragment of the repair factor 53BP1. Cohesin depletion did not affect the local mobility of repair foci in either replicated or unreplicated chromatin. The results indicate that cohesin constrains local spatial dynamics of genomic loci. At the same time, cohesive activity of cohesin is not indispensable for restricting chromatin dynamics, although it enhances the confinement effect. On the other hand, repair foci are less mobile structures than point chromatin loci, and cohesin does not affect their dynamics on the studied time scales. Full article

The Nujiang region of Yunnan is by far the richest tourmaline-producing mining area in China. Since the discovery of the tourmaline-bearing deposit in Yunnan Province in 1980, there have been few comprehensive gemmological studies of this deposit. Therefore, the results of tests on 32 tourmaline samples from the Fugong and Gongshan regions of Yunnan are reported in this paper. The chemical composition of the Yunnan tourmalines was analyzed, and the contents of major trace elements were compared with those of tourmaline samples from different localities reported in the literature to highlight their specific provenance characteristics. Microscopic observation revealed the presence of liquid, gas, and solid inclusions; Raman spectra indicated the presence of constitutional water and CH₄-C₂H₆ dihydrate in the Yunnan tourmalines and also pointed to the influence pattern of the Fe content. The infrared spectrum in the range of 4000–4800 cm⁻¹ showed the frequency of metal cations and hydroxyl groups. Based on the characteristic peaks at 4343 cm⁻¹ and 4600 cm⁻¹, a quick distinction between elbaite and dravite could be made. UV–Vis absorption spectroscopy analysis showed that in yellow tourmalines, Mn²⁺-Ti⁴⁺ IVCT is the main cause of color, while green coloration occurs due to Fe²⁺–Fe³⁺ interactions or Cr³⁺ and V³⁺, and the pink color is caused by Mn³⁺ d-d transitions. The three-dimensional fluorescence spectra revealed the presence of the main fluorescence peaks at λex280/λem320 nm and λex265/λem510 nm in the tourmaline samples analyzed and the fluorescence intensity with Ti and Fe contents. Full article

The fw2.2 (fruit weight 2.2) gene negatively regulates cell division and significantly influences fruit size, but its regulatory mechanisms in pears remain unclear. Here, we investigated how pear FWL (fw2.2-like) genes control cell division using Duli pear, Korla fragrant pear, and Yali pear. During the cell division phase, fluorescence in situ hybridization (FISH) revealed stronger expression of FWL1 and FWL5 in smaller fruits compared to larger ones, with both genes localized in the core and flesh tissues. Gene silencing experiments demonstrated that silencing of FWL5 leads to a significant increase in the number of cells, with a concomitant enlargement of the fruit. Yeast two-hybrid screening identified 147 proteins interacting with FWL5, showing substantial overlap with FWL1 interactors. Key candidates included metallothionein-like protein (MT) and Ole-e-1, with the latter displaying a positive correlation with fruit size during cell division. Bimolecular fluorescence complementation (BiFC) confirmed direct interactions between Ole-e-1 and both FWL1/FWL5. Functional analysis indicated the Ole-e-1 gene family has diverse roles in pear development. We propose that Ole-e-1 interacts with FWL genes to modulate cell division, thereby determining final fruit size. This study uncovers a novel regulatory axis linking cell cycle control and fruit size in pears. Full article