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PubMed1977

Selective inhibition of Bacillus subtilis sporulation by acridine orange and promethazine.

Two structurally similar compounds were found to inhibit sporulation in Bacillus subtilis 168. A dye, acridine orange, and an antischizophrenic drug, promethazine, blocked spore formation at concentrations subinhibitory to vegetative growth, while allowing synthesis of serine protease, antibiotic, and certain catabolite-repressed enzymes. The sporulation process was sensitive to promethazine through T2, whereas acridine orange was inhibitory until T4. The drug-treated cells were able to support the replication of phages phie and phi29, although the lytic cycles were altered slightly. The selective inhibition of sporulation by these compounds may be related to the affinity of some sporulation-specific genes to intercalating compounds.

Acridines↗
PubMed1981

Evaluation of acridine orange stain for detection of Trichomonas vaginalis in vaginal specimens.

Vaginal exudates from 105 patients were examined by direct wet-mount microscopy and acridine orange stain for the presence of Trichomonas vaginalis. Both procedures demonstrated greater than 90% agreement in both sensitivity and specificity. When specimens can be examined immediately after collection, it appears that wet-mount microscopy is almost as sensitive as acridine orange staining for detection of T. vaginalis.

Acridine Orange↗
PubMed1982

Acridine orange staining of Pneumocystis carinii.

Acridine orange was used to stain smears of mouse lung which contained cyst and trophozoite forms of Pneumocystis carinii. Trophozoite forms stained yellow to orange; however, cyst forms did not stain. Acridine orange is a rapid and sensitive method for demonstrating trophozoites of P. carinii in mouse lung tissue.

Acridine Orange↗
PubMed1983

Viability of organisms held in the isolator blood culture system for 15 h and their rapid detection by acridine orange staining.

Sets of three Isolator blood culture tubes were seeded with low numbers of 96 strains of 26 bacterial species (fresh and stock clinical isolates). One tube was processed immediately, and the other two were held at 22 and 34 degrees C for 15 h before processing. Organism recovery was 99, 99, and 98%, respectively. Organism numbers increased at both 22 degrees C (60% of strains) and 34 degrees C (79% of strains). Especially notable was that the increases were seen with most strains of Staphylococcus aureus, streptococcal species, Pseudomonas, and all of the Enterobacteriaceae tested. Seven strains, including Streptococcus pneumoniae and Haemophilus influenzae, although viable, were recovered with a decreased number of organisms at each temperature. Acridine orange staining detected organisms in 53% of those Isolator tubes being held and 71% of those demonstrating a numerical increase, after incubation at 34 degrees C. In addition, it was noted that after processing, 48% of the strains that had increased in number while being held at 34 degrees C resulted in visible growth on agar media in 6 h. The results suggest that up to a 15-h delay in processing Isolator tubes may be possible and that acridine orange staining for the rapid detection of positive cultures may be useful in such a circumstance.

Acridine Orange↗
PubMed1984

Use of Directigen and acridine orange for rapid identification of human blood culture isolates.

Of 7,871 blood cultures from hospital patients, 22 yielded growth of Streptococcus pneumoniae or Haemophilus influenzae type b. The identities of 19 (86%) of these 22 strains could be verified after 18 to 24 h of incubation by application of the Directigen meningitis test kit to the unheated, uncentrifuged supernatant from the cultures; thus, the turnaround time for these cultures was halved. Growth in 16 (72%) of the Directigen-positive cultures was detected by visual inspection, and that in 3 (14%) of the cultures was detected by acridine orange staining. Growth in the three remaining bottles (14%) was detected by blind subculturing after 18 to 24 h or incubation and, therefore, was delayed by 24 h. The systematic application the acridine orange stain was helpful in 40 (44%) of 91 cases for which macroscopic inspection failed to reveal growth after 24 h of incubation.

Acridine Orange↗
PubMed1985

Comparison of the automicrobic system, acridine orange-stained smears, and gram-stained smears in detecting bacteriuria.

We compared the accuracy of the Gram-stained smear, the acridine orange-stained smear, and the AutoMicrobic system (AMS; Vitek Systems, Inc., Hazelwood, Mo.) in screening for bacteriuria, as detected by conventional cultures. For 1,024 clinical specimens, results with the acridine orange-stained smear and the Gram-stained smear were very similar. When read for the presence of one or more microorganisms or leukocytes per 20 oil immersion fields, both smears were highly sensitive (92.1 and 93.3%, respectively) and moderately specific (70.0 and 61.7%, respectively). Sensitivity was greater for specimens yielding greater than or equal to 10(5) CFU/ml (96.1 and 98.9%, respectively) than for those with 10(3) to 10(4) CFU/ml (81.4 and 78.0%, respectively). Preliminary classification based upon the tinctorial and morphological characteristics of the Gram-stained smear was compatible with culture results in nearly all cases. The accuracy of the Gram-stained smears was not influenced by special cleaning of the microscopic slides, or the level of expertise of the microscopist. For 715 specimens, the sensitivity of the AMS in detecting bacteriuria (91.5%) was very similar to that of the stained smears (92.1 and 95.7%, respectively), but the specificity was significantly higher (83.2% versus 42.6 and 70.0%). Detection of microorganisms by the AMS took an average of 6.3 +/- 3.0 h. These data suggest that the Gram-stained smear is easily interpreted, very sensitive, acceptably specific, and still the optimal rapid method for screening for bacteriuria in most clinical microbiology laboratories.

Acridine Orange↗
PubMed1993

Acridine orange staining as a replacement for subculturing of false-positive blood cultures with the BACTEC NR 660.

Despite the customization of growth index thresholds within individual laboratories, use of the BACTEC NR 660 automated blood culture system results in a number of false-positive cultures. The results of Gram staining, acridine orange staining, and subculturing to agar media were evaluated on 210 false-positive blood cultures over a 6-month period. Inclusion of acridine orange staining in the routine workup of false-positive blood cultures can eliminate the need for subculturing.

Acridine Orange↗
PubMed1994

Detection of Bartonella (Rochalimaea) quintana by routine acridine orange staining of broth blood cultures.

Bartonella quintana was isolated from 34 BACTEC nonradiometric aerobic resin blood cultures for 10 adults. Nine patients were initially diagnosed by routine acridine orange staining of routine cultures that had been incubated for 8 days. All subcultures grew on chocolate agar within 3 to 12 days (median, 6 days). The PLUS 26 high-volume aerobic resin medium, combined with acridine orange stain and subculture, is an effective system for detection and isolation of B. quintana from blood.

Acridine Orange↗
PubMed1978

Comparison of pseudorabies virus inactivated by bromo-ethylene-imine, 60Co irradiation, and acridine dye in immune assay systems.

Pseudorabies virus infections among animals, especially swine, have become prevalent in the United States in the past few years. The disease in swine is now economically important. Test systems and antigens are being developed for use in control and disease suppression efforts. Pseudorabies virus was inactivated by three methods: chemically, with bromo-ethylene-imine; physically, with 60Co irradiation; and chemically and physically, with 3,9-diaminoacridine dye followed by exposure to white visible light. The antigenicities of the preparations were determined in the presence of specific antibody in immunodiffusion tests and through immunoelectrophoresis. The latter technique permitted quantitation of either antigen or antibody. In the electrophoretic patterns, the antigenic mass in bromo-ethylene-imine preparations was estimated to be 42 mg/ml, the same as in the untreated control material. After 60Co irradiation, 22 mg/ml was present, in comparison with 50 mg/ml in the untreated control antigen. In contrast, 67 mg/ml was present in the acridine dye-light-treated preparation, in comparison with 58 mg/ml in the untreated control material. A possible explanation for the acridine dye-light-treated preparation values is that photodynamic inactivation interferes with viral maturation during the replicative cycle within cells, with a resulting production of a greater amount of antigen, at least some of which is in the form of defective particles.

Acridines↗
PubMed1967

Acridine sensitivity of bacteriophage T2H in Escherichia coli.

Normally acridine-sensitive, Escherichia coli-T2H complexes are rendered acridine-resistant if the infecting bacteriophage mutant is either pr or q. If these pr or q mutants are treated to produce sensitive revertants, one obtains a mutation at any of several dye-sensitizing (ds) sites in the early enzyme region of the T2 map. The ds mutants are nonspecific suppressors because they reduce the resistance of complexes containing either pr or q to proflavine. The ds mutants are not identical in action, since some make pr or q sensitive to proflavine and quinacrine, and others, to proflavine alone. Two ds mutants have r to r(+) mutation patterns which differ, depending upon whether or not the ds is coupled with r7 (an rII mutant). The mutation patterns of r(+) to r are the same for both ds mutants and for wild type. We suggest that dye sensitization may consist of alterations of early enzymes so as to produce slightly different forms of deoxyribonucleic acid which are in turn dyesensitive.

Acridines↗
PubMed1983

Comparative study of subculture, Gram staining and acridine orange staining for early detection of positive blood cultures.

In view of the importance of a rapid aetiological diagnosis in septicaemia, we compared the results of subculture, Gram staining and acridine orange staining in the detection of positive blood cultures. The study was based on 1013 blood cultures of which 138 were positive by culture. The three techniques were applied 12 h after the specimen was taken in 210 instances, at 24 h in 540 instances and after 48 h in 525. We were able to demonstrate the value of direct examination. Staining with acridine orange yields more positive results than Gram staining and is also simpler.

Acridine Orange↗
PubMed1976

Inheritance of acridine orange R variants in human acrocentric chromosomes.

Sequential Q and acridine orange R banding were performed on 50 normal individuals to determine those individuals with the maximum number of variants by acridine orange reverse banding. In three individuals from different families, one of the two homologs of each of pairs of acrocentric chromosomes was classified as variant. The variant present in two individuals enabled unambiguous determination of the paternal or maternal origin of all 10 acrocentric chromosomes. In the third family, the variants in the third generation were sufficient to determine the origin of homologs in 5 of 10 acrocentric chromosomes. Overall, the parental origin of 36 of 50 acrocentric chromosomes (72%) could be determined. In no case was the pattern of variants found in offsprings and parents incompatible with Mendelian inheritance.

Acridines↗
PubMed1991

Quantitation of early myocardial ischemia using acridine orange fluorescence--an experimental study.

It has been observed in the present investigation that ischemic myocardium consistently produces bright green fluorescence after Acridine Orange (AO) staining. The area of ischemia in the left ventricular myocardium at different time intervals after onset of experimental ischemia has been calculated by use of this AO fluorescence technique. Zonal distribution of ischemia in the epicardial, middle, and endocardial zones has also been evaluated quantitatively from ten minutes to six hours after ligation of the anterior descending branch of the left coronary artery in the Wistar strain of albino rats. No similar study was available for comparison from the literature reviewed. The total area of left ventricular ischemia showed an increase with the prolongation of duration of coronary ligation from 13.39 +/- 2.69 mm2 at ten to twenty-five minutes to 32.99 +/- 5.69 mm2 at six hours after ligation. Statistical analysis of the zonal area of ischemia has shown that ischemia in the middle and endocardial zones was greater than that in the epicardial zone at all time intervals recorded. Middle zone ischemia extended over a larger area than that over the endocardial zone at all intervals except at intervals II (30-45 min), IV (2-2 1/2 hrs) and V (3-4 hrs). The results of this experimental investigation are significant for these point to the value of Acridine Orange fluorescence in detecting early myocardial ischemia and in demarcating zonal differences in ischemia. The authors have successfully utilized the method in a few human cases of clinically suspected myocardial infarction and recommend the technique for routine use to detect early human myocardial ischemia.

Acridine Orange↗
PubMed1976

A phenomenon of photoenhancement of acridine orange-induced fluorescence.

It is a common experience that, with exposure to exciting radiation of the fluorescence microscope, the acridine orange-induced red fluorescence of the nucleus, produced by Feulgen hydrolysis, fades with a concomitant shift to green. The present investigation reports a phenomenon of photoenhancement observed in the hydrolyzed cytoplasm where pale green fluorescence increases in intensity with exposure to exciting radiation. The phenomenon has been noticed in Rhizobium, Oscillatoria, tomato root tip cells and human buccal epithelial cells. It is tentatively concluded that the gain in fluorescence yield is due to certain conformational changes of the acridine orange-protein complex induced by ultraviolet light flux.

Acridines↗
PubMed1979

Mass action and acridine orange staining: static and flow cytofluorometry.

We present results involving an approach to acridine orange staining of intact cells based on basic physicochemical considerations. We show by static microfluorometry of several in vitro and in vivo cell lines that the important parameters for such staining are the molar ratio (Formula: see text), and molar concentration of acridine orange. Differential nuclear DNA and cytoplasmic RNA staining are totally controlled by these two parameters. We show this by a physicochemical model of cell-dye interaction. Finally, we use the method to study the growth parameters of complex in vivo cell populations by automated multiparameter flow microfluorometry. We have explored also, both by static and flow systems, the effect on AO-cell staining of various cell pretreatments such as Triton X-100 and chelating agents.

Acridine Orange↗
PubMed1979

Automatic cell identification and enrichment in lung cancer. II. Acridine orange for cell sorting of sputum.

Fluorescence spectra were obtained from cells from sputum and pleural effusions stained with different fluorescent dyes and fixed by alternate methods. The spectra were referenced to a standard allowing for fluorescence comparisons of unstained and stained cells under various conditions. The metachromasia of acridine orange-stained cells offers nuclear/cytoplasmic differentiation in a single stain; mithramycin and propidium iodide do not. Unstained cells have an appreciable amount of green (546 nm) fluorescence, as does Carbowax in Saccomanno's preservative. Cytoplasm stained with acidine orange also has appreciable green fluorescence. Consequently, cells with much cytoplasm have high total fluorescence. Cytoplasmic fluorescence is negligible with mithramycin or propidium iodide. The metachromasia of acridine orange-stained cells is altered by alcohol and Carbowax levels in fixatives, keeping other factors constant.

Acridine Orange↗
PubMed1989

Acridine derivatives. III. Preparation and antitumor activity of the novel acridinyl-substituted uracils.

In an investigation of a new class of deoxyribonucleic acid (DNA)-intercalating antitumor agents, novel acridinyl-substituted uracils have been synthesized and evaluated for activity against L1210 leukemia in vivo, and against bacteria and fungus. These compounds were prepared by the novel enamine reaction between 9-chloroacridines and 6-aminouracils. The positional effects of substituents on the acridine ring showed that compounds bearing electron-withdrawing groups at the 3- or 6-position of the acridine ring were the most active.

Acridines↗