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PubMedDate not supplied

Validation of the mouse peripheral blood micronucleus assay using acridine orange supravital staining with urethane.

The mouse peripheral blood micronucleus assay using acridine orange supravital staining was compared with the standard bone marrow assay using urethane (ethyl carbamate)-treated mice. Urethane was intraperitoneally injected to CD-1 and BDF1 mice at doses ranging from 62 to 1000 and 62 to 250 mg/kg, respectively. Peripheral blood was collected from the tail 0, 24, 48, and 72 h and bone marrow cells were smeared at 24 and 42 h after the treatment. Although the response of micronucleus induction in peripheral reticulocytes was delayed by about 24 h compared to that in bone marrow polychromatic erythrocytes, the maximum frequencies of micronucleated young erythrocytes were comparable. Therefore, the peripheral blood micronucleus assay using the acridine orange supravital staining method may provide a good alternative to the conventional bone marrow assay.

Acridine Orange↗
PubMedDate not supplied

Antimicrobial activity of phenothiazines, benzo[a]phenothiazines and benz[c]acridines.

The abilities of 14 phenothiazines, 8 benzo[a]phenothiazines and 12 benz[c]acridines to induce an antibacterial effect against Escherichia coli K12 were compared. Several phenothiazines, which showed antiplasmid activity, displayed the most potent antibacterial activity. All benz[c]acridine derivatives were moderately antibacterial, whereas benzo[a]phenothiazines were inactive. The active phenothiazine derivatives had more potent inhibitory activity against fungi, including phytopathogen filamentous, human pathogen filamentous fungi and yeasts, than against gram-positive and -negative bacteria. Taken together with previously reported data, the induction mechanism of antimicrobial and antiplasmid activity by these compounds seems to be different from that of antitumor, differentiation-inducing and carcinogenic activity.

Acridines↗
PubMed2005

Periosteal Ewing's sarcoma treated by photodynamic therapy with acridine orange.

We recently encountered a very rare case of periosteal Ewing's sarcoma (PES), which was treated by surgery followed by photodynamic therapy using acridine orange with radiodynamic therapy. No more than 15 cases of PES have been reported previously in literature. In our case, MRI revealed the tumor to be localized on the cortical surface of the proximal humerus. Histopathological examination of biopsy specimens revealed a small round cell sarcoma suggestive of Ewing's sarcoma or PNET, and immunohistochemistry showed positive staining for MIC2. Although fusion genes EWS-FLI1 and EWS-ERG were undetectable, PES was still considered to be the most likely diagnosis. Therefore, we administered preoperative chemotherapy, as a result of which the tumor shrank to 48% of its original volume. With a view to preserve excellent shoulder and upper limb function, we attempted intralesional tumor resection supported by photodynamic therapy using acridine orange with radiodynamic therapy. After surgery followed by postoperative chemotherapy, the patient has, until the time of writing, had no local tumor recurrence and no evidence of metastatic disease, and can move his shoulder fully and throw a ball well. Since it has been reported that PES has a better prognosis and responsiveness to chemotherapy than intramedullary Ewing's sarcoma, we believe that such reduction surgery with photodynamic therapy might be the strategy of choice to obtain satisfactory limb function in cases of PES.

Acridine Orange↗
PubMedDate not supplied

Antiplasmid activity of phenothiazines, benzo[a]phenothiazines and benz[c]acridines.

Various synthetic derivatives of phenothiazines, benzo[a]phenothiazines and benz[c]acridines were compared for their abilities to induce antiplasmid activity against E. coli F'lac plasmid. Several phenothiazine derivatives were much more potent in antiplasmid activity than benzo[a]phenothiazine- or benz[c]acridine derivatives. Their antiplasmid activity seemed to be enhanced by Cl- or CF3- substitution at 2 C atom, and modified by the side chain length and charge at the L-region of the molecules, as well as by hydrophilicity.

Acridines↗
PubMed2005

[Determination of deoxyribonucleic acid with acridine orange by resonance light scattering technique].

The resonance light scattering (RLS) spectra of acridine orange with deoxyribonucleic acid have been studied. The RLS of acridine orange is greatly enhanced by deoxyribonucleic acid in pH 11. There is a resonance light scattering peak at 324 nm. Under optimal conditions, the linear ranges of the calibration curves are 7.5 x 10(-8) -9.8 x 10(-6) g x mL(-1) for Calf thymus DNA (ct DNA), 3.1 x 10(-8) -7.3 x 10(-6) g x mL(-1) for Fish sperm DNA (fs DNA) with detection limits 20.8 ng x mL(-1) for ct DNA and 20.5 ng x mL(-1) for fs DNA. This method is simple, selectivity and sensitivity has been applied to the determination of deoxyribonucleic acid in mixed samples with satisfactory results.

Acridine Orange↗
PubMedDate not supplied

Acridine orange used for photodynamic therapy accumulates in malignant musculoskeletal tumors depending on pH gradient.

BACKGROUND: Intra-operative photodynamic therapy has been applied with acridine orange (AO-PDT) to human musculoskeletal sarcomas for the past 4 years, resulting in a low local recurrence rate, within 10%, after intra-marginal tumor resection and excellent limb function. However, it is still unclear why acridine orange (AO) specifically accumulates in tumor cells, especially in malignant tumor cells. The purpose of this study was to clarify the mechanism of AO accumulation in malignant musculoskeletal tumors. MATERIALS AND METHODS: Sixty-two musculoskeletal tumors, including 35 malignant and 27 benign tumors, were studied. Using freshly resected tumor material, the extracellular pH (pHe) was measured and the fluorescence intensity of AO accumulated in the tumors was measured by an image analyzer after ex vivo exposure to 1.0 microg/ml AO, followed by blue excitation. In the in vitro study, bafilomycin A1 was exposed to LM 8 mouse osteosarcoma cells, in order to inhibit V-ATPase, subsequently causing a decrease in the pHgradient (deltapH) between the intracellular pH (pHi) and extracellular pH (pHe) or between the pHi and the pHe. AO accumulation and the cytocidal effect of AO were evaluated. RESULTS: The results of the in vivo study, using human materials freshly resected at surgery, revealed that the pHe of the malignant musculoskeletal tumors was significantly lower than that of the benign tumors and normal muscles or adipose tissues and also showed that the AO fluorescence intensity of the malignant musculoskeletal tumors was significantly stronger than that of the benign tumors and normal muscles or adipose tissues. The results also revealed that the AO fluorescence intensity negatively correlated with the pHe in tumors and normal tissues. The in vitro study showed that bafilomycin A1 inhibited the accumulation of AO in acidic organelles, such as lysosomes, and that the cytocidal effect of AO-PDT was also remarkably inhibited. DISCUSSION: Based on results of the in vivo and in vitro studies, it is suggested that malignant musculoskeletal tumors have a large deltapH between the pHi and the pHe or between the pHi and the vaculolar pH and also that a large ApH increases AO accumulation in tumors. We, therefore, believe that AO-PDT may be more effective in highly malignant musculoskeletal tumors than low grade ones, because of their acidity. CONCLUSION: AO accumulation in musculoskeletal tumors was dependent on the ApH between the pHi and the pHe, or between the pHi and the vacuolar pH.

Acridine Orange↗
PubMed2005

Acridine orange staining and viability of the coccoid form of Campylobacter upsaliensis.

Conversion of Campylobacter upsaliensis to the coccoid form during aerobic incubation at 37 degrees C was not prevented by treatment with chloramphenicol and was accompanied by severe decreases in isocitrate dehydrogenase activity and oxygen uptake. Although the coccoid forms fluoresced orange-red by acridine orange staining, agarose gel electrophoresis indicated an extensive degradation of the ribosomal RNA. This suggests that acridine orange staining may not be a good indicator of viability and that the coccoid form of C. upsaliensis at 37 degrees C is degenerative rather than part of the life cycle.

Acridine Orange↗
PubMed1990

Rapid diagnosis of Brugia malayi and Wuchereria bancrofti filariasis by an acridine orange/microhematocrit tube technique.

A microhematocrit tube technique for diagnosis of human filariasis has been previously described. A system incorporating heparin, EDTA, and acridine orange into a microhematocrit tube (Quantitative Blood Count, QBC) has been commercially developed for the quantitation of blood counts and has been used for the diagnosis of malaria. We evaluated this test for its usefulness in the diagnosis of filariasis. Upon centrifugation, the parasites were concentrated in the area of the buffy coat and could be observed through the wall of the tube. The parasites were concentrated further by a plastic float that expands the buffy coat and confines the parasites to the periphery of the tube. Acridine orange stains the DNA of the parasite, and morphologic characteristics can be examined by fluorescence microscopy. The terminal and subterminal nuclei and long cephalic space of Brugia malayi, as well as the short cephalic space and caudal nuclei of Wuchereria bancrofti, were easily recognized and differentiated from each other. Microfilariae were detected in samples diluted to a level of approximately 50/ml.

Acridine Orange↗
PubMed1990

Acridine orange--RNA histofluorescence of sarcomas and small round cell tumors of childhood.

Forty-nine pediatric malignant neoplasms were stained with acridine orange (AO) fluorochrome to qualitatively evaluate cytoplasmic RNA content. The application of AO as a supplementary stain in surgical pathologic diagnosis is based on the premise that specific neoplastic cell types characteristically and consistently contain few or many cytoplasmic ribosomes. Primitive tumors such as Ewing's sarcoma and primitive neuroectodermal tumors showed negative or low-intensity AO-RNA cytoplasmic staining. Differentiated sarcomas such as rhabdomyosarcomas and lymphomas exhibited moderate to strong AO-RNA cytoplasmic fluorescence. Acridine orange--RNA staining provides an easy, convenient, and inexpensive adjunct in the histopathologic differential diagnosis of sarcomas. It is particularly useful for distinguishing Ewing's sarcomas from other small round cell sarcomas of childhood.

Acridine Orange↗
PubMed1990

Acridine orange metachromasia in the cytoplasm of simian rotavirus (SA-11)-infected MA-104 cell cultures.

1. Acridine orange metachromasia was used to determine the distribution of simian rotavirus double-stranded RNA in cultured MA-104 cells 0 to 72 h post-infection. Correlations were made among time of detection and amount of viral antigens, virus yield and the ultrastructural aspects of infected cells. 2. RNAase-resistant cytoplasmic metachromasia appeared 48 h post-infection, 36 h after the initial detection of viral antigens or infectious virions and 24 h after the appearance of the cytopathic effect. 3. Acridine orange staining is thus useful for monitoring the progress of rotaviral infection in cell cultures due to its simplicity and low cost, in spite of its lower sensitivity compared to other techniques evaluated.

Acridine Orange↗
PubMed1991

The differential fluorescence of bacteria stained with acridine orange and the effects of heat.

Some factors affecting the fluorescence of bacteria stained with acridine orange and the direct epifluorescent filter technique (DEFT) were studied. When bacterial cells from a chemostat operated at dilution rates between 0.1 and 0.7/h were used the differential fluorescence observed in the DEFT related to cell 'activity' and the orange fluorescence, which was predominant at high growth rates, may be related to an increase in the RNA content of the cells. Heat affected the colour of cell fluorescence and this was dependent on the cell type and, in particular, age. Uptake of acridine orange into the cells was also found to be an important factor determining the colour of fluorescence. However, with heat-treated cells there was no correlation between the amount of uptake and colour of fluorescence. The relative amounts and degree of denaturation of the different types of nucleic acids remaining in the cells after heat treatment appeared primarily to determine the colour of fluorescence.

Acridine Orange↗
PubMedDate not supplied

Acridine-linked oligonucleotide probes for the short dAT-rich motifs in the 3'-untranslated region of cytokine genes.

We have investigated the use of oligonucleotide probes for identifying cDNA clones containing the short dAT-rich motifs found in the 3'-untranslated region of cytokine genes. To obtain sufficiently stable duplexes between the octameric probes used to identify genes containing the sequence dTATTTATT and its complement, it was necessary to couple an intercalating agent, an acridine derivative (acr), to the 5'-positions of the probes. The resulting octamers 5'-acr-dAATAAATA and, particularly, 5'-acr-dTATTTATT were successfully used to distinguish the complementary sequences in cDNA from internal, single point mismatched sequences. Southern blot analyses of plasmids containing IL-1 beta and IL-8 gave positive results with the 3' degenerate probe, 5'-acr-dTATTTATTN, clearly showing that the very short probe approach can be used in this type of analysis. Subsequently, in slot blot analyses we found that, even without the degenerate nucleotide, N, plasmids bearing cytokine sequences with at least 7 contiguous matched nucleotides could be unambiguously identified with 5'-acr-dTATTTATT. Unfortunately, because of the ubiquity of these dAT-rich sequences in bacterial DNA, it was not possible to use these probes for direct colony screening. In contrast to the results obtained with DNA, at the RNA level, with IL-1 beta mRNA bound to nitrocellulose, the hybrid formed with 5'-acr-dAATAAATA was very unstable, even in 1M LiCl solution at 2 degrees C; however, in the same salt solution the slightly longer acridine-coupled probes 5'-acr-dAATAAATAGGG and 5'-acr-dAAAGAACAA remained hybridized to their complementary sequences up to about 18 degrees C.

Acridines↗
PubMed1985

[Acridine orange: a fluorochrome of nucleic acids for the study of muscle and nerve cells].

Acridine orange renders nucleic acids fluorescent when forming complexes with them: D.N.A. is coloured luminous yellow while R.N.A. is a brilliant orange. Its application to sections of muscle biopsies permits the identification of certain muscle fibres, recognized by their bright orange fluorescence which contrasts with the pale green of mature, normal myofibrils in three specific situations: denervation, regeneration following injury, and the rapid growth and maturation of fetal muscle. Moreover, myopathic nonregenerative atrophy, as well as histochemical type-specific atrophies, do not exhibit this same orange fluorescence. Because neurons, particularly motor neurons, contain large amounts of R.N.A. (by contrast with glial cells which have very little) the application of acridine orange in the central nervous system might help in understanding the development of the fetal brain, the evolution of some neurologic diseases, and the maturation of brain tumors. Since the technique does not require the exclusive use of frozen tissues, but also may be applied to formalin-fixed tissue in paraffin sections, it is well adapted to the study of tissue obtained at autopsy.

Acridine Orange↗
PubMed1986

A study of osteoclasts on calvaria of normal and osteopetrotic (mi/mi) mice by vital staining with acridine orange.

A novel staining procedure for enumerating osteoclasts on neonatal mouse calvaria with the vital fluorescent dye acridine orange is described. It has the advantage over Barnicot's neutral-red method in that the nuclei and cytoplasm of the osteoclast are stained differentially. The osteopetrotic calvarium (mi/mi) has fewer multinucleate osteoclasts than its normal counterpart (mi/+) and they are differently distributed. The osteopetrotic calvarium has more mononucleate cells which stain like osteoclasts with acridine orange than the normal calvarium and these cells also are differently distributed. These mononuclear cells may be mononuclear osteoclasts or their precursors. These observations suggest that the defect resulting in this osteopetrosis lies with osteoclast differentiation.

Acridine Orange↗
PubMed1988

[A method of bacterial count by epifluorescence with acridine orange. Application to skin biopsies performed in burnt patients].

Acridine orange was used for staining and counting micro-organisms obtained from 136 skin biopsies performed in burned patients. The number of organisms per gram of tissue was compared to the number of colony-forming-units (CFU) calculated from cultures of the same biopsies. The staining method was positive in 97 per cent of septic samples, and in 25 per cent of these it proved more sensitive than bacterial cultures, with a 100 to 1000-fold greater number of pathogens detected. Acridine orange also demonstrated bacteria in 69 biopsies which remained sterile after culture. In some cases, the same bacterial species was found in other samples taken a few days later.

Acridine Orange↗
PubMed1989

Detection of microorganisms in clinical specimens using slides prestained with acridine orange (AOS).

Slides prestained with acridine orange (AOS) were used for direct examination of microorganisms in clinical specimens. The method is simpler than a commonly used acridine orange stain (AO) and has the advantage of being a supravital stain. Results indicate that the AOS method is sensitive, and allows rapid detection of microorganisms, even in the specimens containing few microorganisms, e.g. cerebrospinal fluid. Moreover AOS may be the choice method for detection of Trichomonas vaginalis.

Acridine Orange↗
PubMed1989

[Distribution of RNA during neuronal migration in developing normal and dysplastic brains in man. Acridine orange study].

Acridine orange, a fluorochrome stain of nucleic acids, was used to study neuronal maturation in human brains during development. The central nervous system in 15 normal fetuses and neonates of 16 to 44 weeks gestational age were examined, as well as 24 dysplastic brains exhibiting abnormal cellular migrations. The increase in cytoplasmic RNA of neurons coincides with the onset of neurotransmitter synthesis, temporally synchronized to begin after migration in the cerebral hemispheres but before migration of the external granular layer of the cerebellum. The presence or absence of orange fluorescence in heterotopic nerve cells serves as a marker of the state of maturity by indicating whether the cell was still migrating at the time of death or had already arrived at its definitive site in the brain, whether normal or abnormal. Furthermore, the type of neurotransmitter produced by a given neuron influences ribosomal concentration and hence the intensity of orange RNA fluorescence. Neurons that secrete peptides, acetylcholine, or monoamines show stronger fluorescence than do those synthesizing simple amino acids as transmitters. Transitory transmitters of developing brain may account for stronger fluorescence in certain neurons of the fetus than in the adult, an example being the granule cells of the cerebellar cortex. Acridine orange is a useful supplementary tool in pediatric neuropathology.

Acridine Orange↗
PubMed1989

[Spinal muscular atrophy: disappearance of RNA fluorescence of degenerating motor neurons. An acridine orange study].

The histochemical distribution of nucleic acids has been studied in degenerating motor neurons of 9 children who died with spinal muscular atrophy, using the fluorochrome acridine orange. Ribonucleic acid (RNA) fluorescence disappeared abruptly from involved motor neurons without chromatolysis, attenuation of intensity, or other intermediate transitions that follow axotomy or hypoxic insults. We found a nearly identical pattern in 3 adults with amyotrophic lateral sclerosis. The findings in older subjects are complicated, however, by the presence of cytoplasmic lipofuscin. The autofluorescence of this pigment is inhibited by acridine orange. Our results support the hypothesis that spinal muscular atrophy is a disturbance of the genetically coded mechanism that arrests the programmed physiological death of surplus motor neuroblasts after a certain time in embryonic life, so that the normal lethal developmental process becomes pathological by persisting postnatally. A failure of RNA transcription seems to be primary and results in a failure of synthesis of neurotransmitters, of enzymes, and of cytoplasmic proteins. Consequent inanition leads to cell death.

Acridine Orange↗