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Cytometry. Part a : the Journal of the... Jun 2020Briefly depicted are the publications in CYTOMETRY that received the highest frequency of citations. Among them are seminal papers describing application of...
Briefly depicted are the publications in CYTOMETRY that received the highest frequency of citations. Among them are seminal papers describing application of metachromatic fluorochrome acridine orange to differentially stain DNA versus RNA or to analyze susceptibility of DNA in situ to denaturation; both features being markers of different sections of the cell cycle including identification of noncycling quiescent cells. The papers reviewing detection of cyclins D1, E, A or B1, each in relation to cell cycle phase, were also among the highly cited ones. The highest citation rates received publications describing development of the TUNEL methodology to detect apoptotic DNA fragmentation, and more recently expression of ϒH2AX to reveal DNA damage. © 2020 International Society for Advancement of Cytometry.
Topics: Acridine Orange; Cell Cycle; DNA; Flow Cytometry; Fluorescent Dyes
PubMed: 32511890
DOI: 10.1002/cyto.a.24043 -
Cancers Jul 2021In bone sarcomas, extracellular proton accumulation is an intrinsic driver of malignancy. Extracellular acidosis increases stemness, invasion, angiogenesis, metastasis,... (Review)
Review
In bone sarcomas, extracellular proton accumulation is an intrinsic driver of malignancy. Extracellular acidosis increases stemness, invasion, angiogenesis, metastasis, and resistance to therapy of cancer cells. It reprograms tumour-associated stroma into a protumour phenotype through the release of inflammatory cytokines. It affects bone homeostasis, as extracellular proton accumulation is perceived by acid-sensing ion channels located at the cell membrane of normal bone cells. In bone, acidosis results from the altered glycolytic metabolism of bone cancer cells and the resorption activity of tumour-induced osteoclasts that share the same ecosystem. Proton extrusion activity is mediated by extruders and transporters located at the cell membrane of normal and transformed cells, including vacuolar ATPase and carbonic anhydrase IX, or by the release of highly acidic lysosomes by exocytosis. To date, a number of investigations have focused on the effects of acidosis and its inhibition in bone sarcomas, including studies evaluating the use of photodynamic therapy. In this review, we will discuss the current status of all findings on extracellular acidosis in bone sarcomas, with a specific focus on the characteristics of the bone microenvironment and the acid-targeting therapeutic approaches that are currently being evaluated.
PubMed: 34359749
DOI: 10.3390/cancers13153848 -
Frontiers in Oncology 2019Acridine orange (AO) was first extracted from coal tar in the late nineteenth century and was used as a fluorescent dye. In this paper, we review emergent research... (Review)
Review
Acridine orange (AO) was first extracted from coal tar in the late nineteenth century and was used as a fluorescent dye. In this paper, we review emergent research about novel applications of AO for fluorescence surgery and cancer therapy. We performed a systematic search in the MEDLINE, PubMed, Cochrane library, Google Scholar, Embase, Web of Science, and Scopus database using combinations of the term "acridine orange" with the following: "surgical oncology," "neuropathology," "microsurgery," "intraoperative fluorescence," "confocal microscopy," "pathology," "endomicroscopy," "guidance," "fluorescence guidance," "oncology," "surgery," "neurooncology," and "photodynamic therapy." Peer-reviewed articles published in English were included in this review. We have also scanned references for relevant articles. We have reviewed studies on the various application of AO in microscopy, endomicroscopy, intraoperative fluorescence guidance, photodynamic therapy, sonodynamic therapy, radiodynamic therapy. Although the number of studies on the clinical use of AO is limited, pilot studies have demonstrated the safety and feasibility of its application as an intraoperative fluorescent dye and as a novel photo- and radio-sensitizator. Further clinical studies are necessary to more definitively assess the clinical benefit AO-based fluorescence guidance, therapy for sarcomas, and to establish feasibility of this new approach for the treatment of other tumor types.
PubMed: 31612102
DOI: 10.3389/fonc.2019.00925 -
Biochimica Et Biophysica Acta.... Jun 2023The efficiency of methylene blue (MB) and acridine orange (AO) for photodynamic therapy (PDT) is increased if encapsulated in liposomes. In this paper we determine the...
The efficiency of methylene blue (MB) and acridine orange (AO) for photodynamic therapy (PDT) is increased if encapsulated in liposomes. In this paper we determine the molecular-level interactions between MB or AO and mixed monolayers of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dipalmitoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (DPPG) and cholesterol (CHOL) using surface pressure isotherms and polarization-modulated infrared reflection absorption spectroscopy (PM-IRRAS). To increase liposome stability, the effects from adding the surfactants Span® 80 and sodium cholate were also studied. Both MB and AO induce an expansion in the mixed monolayer, but this expansion is less significant in the presence of either Span® 80 or sodium cholate. The action of AO and MB occurred via coupling with phosphate groups of DPPC or DPPG. However, the levels of chain ordering and hydration of carbonyl and phosphate in headgroups depended on the photosensitizer and on the presence of Span® 80 or sodium cholate. From the PM-IRRAS spectra, we inferred that incorporation of MB and AO increased hydration of the monolayer headgroup, except for the case of the monolayer containing sodium cholate. This variability in behaviour offers an opportunity to tune the incorporation of AO and MB into liposomes which could be exploited in the release necessary for PDT.
Topics: Methylene Blue; Acridine Orange; Liposomes; Sodium Cholate; Spectrophotometry, Infrared
PubMed: 37031871
DOI: 10.1016/j.bbamem.2023.184156 -
Analytical Biochemistry Dec 2022Eukaryotic cells are compartmentalized into membrane-bound organelles, allowing each organelle to maintain the specialized conditions needed for their specific...
Eukaryotic cells are compartmentalized into membrane-bound organelles, allowing each organelle to maintain the specialized conditions needed for their specific functions. One of the features that change between organelles is lumenal pH. In the endocytic and secretory pathways, lumenal pH is controlled by isoforms and concentration of the vacuolar-type H-ATPase (V-ATPase). In the endolysosomal pathway, copies of complete V-ATPase complexes accumulate as membranes mature from early endosomes to late endosomes and lysosomes. Thus, each compartment becomes more acidic as maturation proceeds. Lysosome acidification is essential for the breakdown of macromolecules delivered from endosomes as well as cargo from different autophagic pathways, and dysregulation of this process is linked to various diseases. Thus, it is important to understand the regulation of the V-ATPase. Here we describe a high-throughput method for screening inhibitors/activators of V-ATPase activity using Acridine Orange (AO) as a fluorescent reporter for acidified yeast vacuolar lysosomes. Through this method, the acidification of purified vacuoles can be measured in real-time in half-volume 96-well plates or a larger 384-well format. This not only reduces the cost of expensive low abundance reagents, but it drastically reduces the time needed to measure individual conditions in large volume cuvettes.
Topics: Acridine Orange; Vacuolar Proton-Translocating ATPases; Vacuoles; Endosomes; Saccharomyces cerevisiae; Lysosomes; Hydrogen-Ion Concentration
PubMed: 36167157
DOI: 10.1016/j.ab.2022.114927 -
Colloids and Surfaces. B, Biointerfaces Dec 2022Photodynamic therapy uses photosensitizer molecules for the photo-mediated treatment of several diseases such as cancer and skin disorders. However, most of the...
Photodynamic therapy uses photosensitizer molecules for the photo-mediated treatment of several diseases such as cancer and skin disorders. However, most of the photosensitizer molecules present problems such as aggregation and low solubility in physiological environments which hinders the treatment efficacy. To overcome these problems, the development of stable liposomes loading photosensitizing molecules as delivery systems can be explored as promising alternatives to enhance cellular uptake and the therapy's efficacy. In this work, liposomes composed by different lipids with or without surfactants were characterized for the encapsulation of photosensitizer molecules such as Methylene Blue (MB) and Acridine Orange (AO). Liposomes were produced by the thin-film hydration method followed by extrusion to reduce particle size and were characterized by Dynamic Light Scattering and Atomic Force Microscopy. Encapsulation efficiency was evaluated as well as the release profile of these molecules from the liposome systems. Cytotoxicity and phototoxicity studies were performed on keratinocytes with and without carcinoma. Results showed that liposome's stability depends on the composition of lipids regardless of the presence of surfactants. Most stable liposomes were those with cholesterol plus the surfactants Span® 80 or sodium cholate that were able to provide higher stability for the liposomes considering the MB and AO encapsulation. Encapsulation efficiency (EE) studies revealed that AO had greater affinity for the vesicles presenting high EE (>98%) while for MB the encapsulation was, in general, moderate (between 63% and 86%). Greater phototoxicity was observed for MET1 squamous cell carcinoma (SCC) cells treated with AO liposomes, achieving similar half-maximal inhibition concentration (IC) as for the free drug. Finally, two different possible approaches were found, namely, MB-liposomes with potential as a cytotoxic agent for cancer cells; and AO liposomes with a great phototoxicity potential at very low concentrations.
Topics: Humans; Liposomes; Acridine Orange; Photosensitizing Agents; Methylene Blue; Photochemotherapy; Skin Neoplasms; Surface-Active Agents; Lipids
PubMed: 36215895
DOI: 10.1016/j.colsurfb.2022.112901 -
Anticancer Research Nov 2022This study evaluated the effect of haematogenous administration of acridine orange (AO) alone and in combination with zoledronate (ZOL) on bone metastases.
BACKGROUND/AIM
This study evaluated the effect of haematogenous administration of acridine orange (AO) alone and in combination with zoledronate (ZOL) on bone metastases.
MATERIALS AND METHODS
E0771 cells (1.0×10 cells/10 μl) were injected directly into the right femur of female mice. The mice were divided into five groups according to treatment (drugs and irradiation) and were reared and sacrificed after 6 weeks. Micro-computed tomography (μCT) was performed to calculate the destruction rate of the femur bone. We measured tumour weight and volume at sacrifice and performed terminal deoxynucleotidyl transferase dUTP Nick-End Labelling staining of tumours.
RESULTS
At 4 weeks, the bone destruction rate was lower in the AO+ZOL group than in the radiation group. At 6 weeks, the AO+ZOL group had a lower bone destruction rate than the control and radiation groups; the ZOL group had a lower rate than the radiation group. The AO and AO+ZOL groups had suppressed tumour weight and volume compared to the control and radiation groups. The number of extraosseous apoptotic cells was higher in the AO+ZOL group than in all other groups except the AO group.
CONCLUSION
In a model of local bone metastasis of breast cancer, haematogenous administration of AO reduced tumour size and more so when combined with ZOL.
Topics: Animals; Female; Mice; Acridine Orange; Bone Density Conservation Agents; Bone Neoplasms; Diphosphonates; DNA Nucleotidylexotransferase; Imidazoles; Osteolysis; X-Ray Microtomography; Zoledronic Acid; Breast Neoplasms
PubMed: 36288846
DOI: 10.21873/anticanres.16041 -
International Journal of Molecular... Jan 2023Swim-up selected human sperm were incubated with 7 ng F-neuroprostanes (F-NeuroPs) for 2 and 4 h. Sperm motility and membrane mitochondrial potential (MMP) were...
Swim-up selected human sperm were incubated with 7 ng F-neuroprostanes (F-NeuroPs) for 2 and 4 h. Sperm motility and membrane mitochondrial potential (MMP) were evaluated. The percentage of reacted acrosome was assessed by pisum sativum agglutinin (PSA). Chromatin integrity was detected using the acridine orange (AO) assay and localization of the ryanodine receptor was performed by immunofluorescence analysis. Sperm progressive motility ( = 0.02) and the percentage of sperm showing a strong MMP signal ( = 0.012) significantly increased after 2 h F-NeuroP incubation compared to control samples. The AO assay did not show differences in the percentage of sperm with dsDNA between treated or control samples. Meanwhile, a significantly higher number of sperm with reacted acrosomes was highlighted by PSA localization after 4 h F-NeuroP incubation. Finally, using an anti-ryanodine antibody, the immunofluorescence signal was differentially distributed at 2 and 4 h: a strong signal was evident in the midpiece and postacrosomal sheath (70% of sperm) at 2 h, whereas a dotted one appeared at 4 h (53% of sperm). A defined concentration of F-NeuroPs in seminal fluid may induce sperm capacitation via channel ions present in sperm cells, representing an aid during in vitro sperm preparation that may increase the positive outcome of assisted fertilization.
Topics: Humans; Male; Neuroprostanes; Sperm Motility; Seeds; Spermatozoa; Acrosome; Acridine Orange
PubMed: 36674450
DOI: 10.3390/ijms24020935 -
International Journal of Molecular... Oct 2022Diflubenzuron is an insecticide that serves as a chitin inhibitor to restrict the growth of many harmful larvae, including mosquito larvae, cotton bollworm and flies....
Diflubenzuron is an insecticide that serves as a chitin inhibitor to restrict the growth of many harmful larvae, including mosquito larvae, cotton bollworm and flies. The residue of diflubenzuron is often detected in aquaculture, but its potential toxicity to aquatic organisms is still obscure. In this study, zebrafish embryos (from 6 h to 96 h post-fertilization, hpf) were exposed to different concentrations of diflubenzuron (0, 0.5, 1.5, 2.5, 3.5 and 4.5 mg/L), and the morphologic changes, mortality rate, hatchability rate and average heart rate were calculated. Diflubenzuron exposure increased the distance between the venous sinus and bulbar artery (SV-BA), inhibited proliferation of myocardial cells and damaged vascular development. In addition, diflubenzuron exposure also induced contents of reactive oxygen species (ROS) and malondialdehyde (MDA) and inhibited the activity of antioxidants, including SOD (superoxide dismutase) and CAT (catalase). Moreover, acridine orange (AO) staining showed that diflubenzuron exposure increased the apoptotic cells in the heart. Q-PCR also indicated that diflubenzuron exposure promoted the expression of apoptosis-related genes (bax, bcl2, p53, caspase3 and caspase9). However, the expression of some heart-related genes were inhibited. The oxidative stress-induced apoptosis damaged the cardiac development of zebrafish embryos. Therefore, diflubenzuron exposure induced severe cardiotoxicity in zebrafish embryos. The results contribute to a more comprehensive understanding of the safety use of diflubenzuron.
Topics: Acridine Orange; Animals; Antioxidants; Cardiotoxicity; Catalase; Chitin; Diflubenzuron; Embryo, Nonmammalian; Insecticides; Malondialdehyde; Oxidative Stress; Reactive Oxygen Species; Superoxide Dismutase; Tumor Suppressor Protein p53; Water Pollutants, Chemical; Zebrafish; bcl-2-Associated X Protein
PubMed: 36233243
DOI: 10.3390/ijms231911932