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PubMed1977

Influence of phenobarbital pretreatment on biliary rose bengal excretion in rats.

Plasma concentration, hepatic uptake and biliary excretion of intravenously administered rose bengal was determined in rats pretreated with phenobarbital (50 mg/kg) daily, for four days). After an initial (0--16 min) rapid fall in plasma rose bengal concentration caused by hepatic uptake of the dye, the curves in control and pretreated rats did not differ from each other either after administration of a small (5 mg/kg) or a large (50 mg/kg) dose. Hepatic rose bengal concentration was significantly lower in pretreated animals than in the control group. Since liver weight was higher in the phenobarbital pretreated animals than in the controls, the total amount of rose bengal taken up by the liver did not differ in the two groups. The biliary excretion of low dose (5 mg/kg) rose bengal was significantly higher in phenobarbital pretreated than in the control rats but with doses of 50 mg/kg and 100 mg/kg no difference was observed. These doses of rose bengal diminished the increased bile flow caused by phenobarbital.

Animals↗
PubMed1989

Singlet oxygen-induced arrhythmias. Dose- and light-response studies for photoactivation of rose bengal in the rat heart.

In a study of aerobically perfused rat hearts, the in situ photoactivation (530-590 nm) of rose bengal (a process that leads to the production of singlet oxygen and superoxide) has been shown to lead to the rapid development of electrocardiographic abnormalities and arrhythmias. With rose bengal concentrations of 1,000, 500, 250, 100, and 50 nmol/l (n = 6/group), photoactivation (3,600 lx) led to electrocardiographic changes (inversion of the T wave, Q-T prolongation, or both) after 3.8 +/- 0.9, 4.5 +/- 0.7, 11.8 +/- 2.1, 24.8 +/- 3.9, and 65.3 +/- 6.0 seconds), respectively; ventricular premature beats occurred in 100% of hearts after 0.5 +/- 0.2, 1.1 +/- 0.3, 2.2 +/- 0.7, 4.4 +/- 0.8, and 6.6 +/- 1.2 minutes, respectively. Ventricular tachycardia occurred in 83%, 83%, 83%, 67%, and 50% of hearts after 2.1 +/- 0.2, 2.1 +/- 0.4, 2.8 +/- 0.7, 5.7 +/- 2.0, and 11.2 +/- 1.9 minutes, respectively, and complete atrioventricular block in 100%, 100%, 100%, 100%, and 67% of hearts after 3.8 +/- 0.7, 6.5 +/- 1.0, 5.5 +/- 0.9, 13.8 +/- 1.0, and 14.1 +/- 0.9 minutes, respectively. With a fixed concentration (250 nmol/l) of rose bengal, similar light-response relations were observed. Photoactivation of rose bengal had no effect on heart rate but caused a transient (0-4 minutes) vasodilation followed by a progressive vasoconstriction. In further studies in which rose bengal was washed out for 10 minutes before photoactivation, several arrhythmias still developed, indicating that rose bengal binds strongly to tissue and acts as a cellular level rather than in the vascular compartment. To assess the reversibility of rose bengal-induced effects, hearts (n = 6/group) were perfused with rose bengal (250 nmol/l) for 1, 2, 4, 6, and 20 minutes followed by perfusion in the dark for 19, 18, 16, 14, and 0 minutes, respectively. During dark perfusion, the incidence of arrhythmias declined and any decrease in coronary flow was reversed. However, analysis of contents of adenosine triphosphate, creatine phosphate, lactate, and creatine kinase leakage indicated the occurrence of severe injury that did not abate on termination of photoactivation. Finally, although many arrhythmias developed before the onset of vasoconstriction, the reduction in flow with consequent ischemia was shown to exacerbate vulnerability to arrhythmias. In conclusion, short-lived reactive oxygen intermediates such as singlet oxygen and superoxide, which are produced during the photoactivation of rose bengal, can cause rapid and major damage to the heart and its function.

Animals↗
PubMed1994

Characterization of photodynamic actions of rose bengal on cultured cells.

PURPOSE: The authors have previously reported successful photodynamic occlusion of corneal blood vessels using intravenous rose bengal and argon green laser irradiation. To explore the action mechanism of this novel technique of photothrombosis, they examined the photodynamic effect of rose bengal on cultured fibroblasts, smooth muscle cells, and vascular endothelium--the cellular components of blood vessels. METHODS: Five types of cells were exposed to different concentrations of rose bengal and argon green laser irradiation. The irradiated cell areas were analyzed by fluorescence microscopy and fluorometry. Various potential quenchers and proteins were tested for their modulation of the photodynamic action. RESULTS: Upon irradiation with 16 W/cm2 of argon green laser light in conjunction with rose bengal concentrations extending above 1 x 10(-4) M, all cultured cell types showed a dose-dependent photobiologic effect characterized by constriction and detachment of the laser-irradiated cell region from the rest of the cell monolayer. In addition, there was dye photobleaching and development of a blue shift of the fluorescence excitation and emission maxima in the irradiated cell areas. Binding of rose bengal to intracellular components was demonstrated by fluorescence microscopy and by fluorometry showing a red shift of the excitation maximum compared to the maximum in solution. This binding was a prerequisite for expression of the described photobiologic effect, because polymer-conjugated rose bengal (Sensitox II) failed to reproduce it. The addition of native or heat-inactivated bovine serum albumin or catalase decreased this photobiologic effect also owing to dye binding, as indicated by G-75 Sephadex gel filtration chromatography. CONCLUSION: These results indicate that the specific photobiologic effect of monolayer contraction, which simulates the vasoconstriction seen during photothrombosis under argon green irradiation, appears to be caused by the photochemical interaction of rose bengal bound with intracellular components.

Animals↗
PubMed1994

Efficacy of different Rose Bengal and complement fixation antigens for the diagnosis of Brucella melitensis infection in sheep and goats.

Fifteen different Rose Bengal antigens showed large differences with respect to pH, cell concentration and agglutination with the international standard anti-Brucella abortus serum, demonstrating the lack of international standardisation. Their sensitivity and specificity, compared with that of the complement fixation test, were evaluated for the diagnosis of B melitensis infection in culture-positive sheep, brucella-free ewes, and sheep and goats belonging to field flocks under different epidemiological conditions. All the Rose Bengal antigens and the complement fixation test had 100 per cent specificity when testing brucella-free sheep or animals belonging to flocks in unvaccinated brucellosis-free areas, but there were large differences in sensitivity between the Rose Bengal antigens with sera from culture-positive sheep or from animals belonging to infected flocks. When using the most sensitive antigen, no difference was observed in Rose Bengal sensitivity between animals infected with either biovar 1 or biovar 3 of B melitensis. The relationship between the sensitivity of the Rose Bengal antigens and cell concentration was unclear, but their sensitivity was related to the standardisation of the antigens with the international standard serum. The complement fixation test was less sensitive than the Rose Bengal test when testing culture-positive sheep. When testing sera from animals belonging to infected flocks with antigens standardised according to European Union rules, no great differences were observed in the sensitivities of the two tests. However, great differences in sensitivity between the Rose Bengal antigens were observed with sera from animals belonging to flocks with low levels of prevalence.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1982

Comparison of the photodynamic action of Rose Bengal and tetrachlorosalicylanilide on isolated porcine erythrocyte membranes.

The light-mediated effects of Rose Bengal and 3,3',4',5-tetrachlorosalicylanilide (T4CS) on porcine erythrocyte membranes have been investigated. Irradiation in the presence of Rose Bengal and oxygen causes extensive destruction of the unsaturated fatty acids of the erythrocyte membrane. This results in a decrease in the membrane flexibility as measured by a nitroxide spin probe. Irradiation in the presence of T4CS and oxygen had no measurable effect on the unsaturated fatty acids or on the membrane flexibility. Irradiation of erythrocyte membranes both in the presence of Rose Bengal and oxygen and of T4CS gave rise to polymerisation of the membrane proteins. This was evident by polyacrylamide gel electrophoresis and by freeze-fracture electron microscopy. Aggregation of membrane proteins could be observed with low levels of Rose Bengal and short irradiation times at which no loss of unsaturated fatty acids could be detected. Irradiation of the n-butanol-extracted apoprotein of porcine erythrocyte membranes both in the presence of Rose Bengal and of T4CS resulted in polymerisation of the protein as measured by gel electrophoresis and electron microscopy. The results obtained from the two photodynamic compounds are compared in terms of their different mechanisms of action on the membrane. The implications of the results in determining the molecular events leading to photohaemolysis are discussed.

Allergens↗
PubMed1987

[Use of the rose bengal test in the diagnosis of brucellosis in hares].

The diagnostic effectiveness of the Rose bengal test was verified in a set of 72 samples of the blood of hare shot in a focus of brucellosis; the blood was found to be serologically positive by the following methods: rapid agglutination with serum, surface fixation test, method of slow agglutination, and the complement fixation reaction. The Rose bengal test was found positive in 57 cases (79.17%), the surface agglutination test was positive in 60 cases (83.33%), rapid agglutination with serum in 41 cases (56.94%), slow agglutination in 55 cases (76.39%), and the complement-fixation reaction in 63 cases (87.50%). In nine cases (12.5%) a positive reaction was obtained only when the Rose bengal test was used. The results confirm that the Rose bengal test is a sensitive qualitative serological method of low technical and time requirements, suitable for routine diagnostic uses. In the field epizootological investigations in the foci of hare brucellosis, the Rose bengal test is a good screening serological method.

Agglutination Tests↗
PubMed1998

Toxic effect of rose bengal dye on the living human corneal epithelium.

PURPOSE: To investigate whether rose bengal dye at a concentration of 1% has a toxic in vivo effect on the human corneal epithelium. MATERIAL AND METHODS: 80 patients with presumed keratoconjunctivitis sicca (KCS) were examined with break-up time, Schirmer 1 test, and staining with fluorescein sodium and rose bengal. The corneal surface was observed in the slit lamp and photographed by means of non-contact photomicrography. RESULTS: After the application of rose bengal 1% in eyes preinstilled with fluorescein sodium 1% myriads of green dots appeared on the corneal surface in all 32 patients with KCS according to Copenhagen criteria, in 17/18 with low Schirmer 1 test values only, and in 21 of the remaining 30 in whom the KCS diagnosis wasn't verified. CONCLUSIONS: The appearance of green microdots after the application of rose bengal was probably due to the interruption of intercellular junctions and the penetration of fluorescein stained fluid below the superficial cells. Whether this phenomenon reveals an unhealthy corneal surface and thus has a clinical significance has to be further investigated.

Adult↗
PubMed1984

Cornea endothelial rose bengal photosensitization. Effect on permeability, sodium flux, and ultrastructure.

Rabbit corneal endothelial cells demonstrated an increase of endothelial membrane permeability to inulin following perfusion with 5 X 10(-6) M rose bengal and exposure to 1050 muW/cm2 incandescent light for either 1 min or 5 min when compared to corneas similarly perfused with rose bengal but not exposed to light. Endothelial permeability to dextran was unaltered following the photosensitization reaction. Rose bengal perfusion in the absence of light caused an increase in the unidirectional sodium fluxes (J stroma endothelium and J endothelium stroma); however, a net sodium flux was still present and directed from the stroma to the endothelium. Exposure of rose bengal perfused endothelial cells to incandescent light resulted in a further increase in the unidirectional passive sodium flux in both directions and a reduction in the active component of the sodium flux from stroma to endothelium. This caused a reduction in the net active transport of sodium across the endothelium following the photosensitization reaction. Transmission electron microscopy showed no alteration of endothelial cell ultrastructural integrity following rose bengal perfusion in the absence of light. However, rose bengal perfusion accompanied by exposure to 1050 muW/cm2 incandescent light caused swelling of endoplasmic reticulum and mitochondria. The data indicate that rose bengal photosensitization has an adverse effect on endothelial cell energy production as well as transport and barrier function.

Animals↗
PubMed2006

Rose bengal and lissamine green inhibit detection of herpes simplex virus by PCR.

PURPOSE: To determine whether rose bengal and lissamine green affect polymerase chain reaction (PCR) detection of herpes simplex virus (HSV). DESIGN: Laboratory investigation. METHODS: Diagnostic corneal scrapings were evaluated for PCR inhibitory activity. Dacron swabs inoculated with rose bengal and lissamine green were processed as clinical samples, inoculated with control HSV, varicella zoster (VZV), cytomegalovirus (CMV), and toxoplasma DNA and prepared for PCR. The effects of calcium alginate and cotton swabs were also evaluated. RESULTS: Rose bengal, lissamine green, and calcium alginate not only inhibit PCR detection of HSV DNA, but also detection of VZV, CMV, and toxoplasma DNA. This inhibition could be overcome by serial dilution and by DNA purification of the sample before PCR. CONCLUSIONS: Rose bengal, lissamine green, and calcium alginate can inhibit PCR detection of HSV DNA. Clinical scrapings to be sent for PCR diagnostic testing should be taken before instillation of rose bengal or lissamine green.

Alginates↗
PubMed1989

Rose bengal score--a possible key parameter when evaluating disease level and progression in primary Sjögren's syndrome.

The rose bengal score is one of the most commonly used tests for evaluation of ocular surface epithelial damage. The test is used in most Sjögren's syndrome criteria. We examined 24 female and four male patients with primary Sjögren's syndrome (primary SS) in order to evaluate possible correlation between the various tests for keratoconjuncivitis sicca (KCS), and for possible correlations to xerostomia and p-IgG levels. Among the KCS tests a high rose bengal score appeared to be the key parameter, being correlated to low break-up time (P less than 0.01), low tear lysozyme (P less than 0.01), appearance of snake-like chromatin in conjunctival imprints (P less than 0.05), low sialometry (P greater than 0.01) and high p-IgG (P less than 0.01). We followed another group of patients with primary SS (30 females and four males) for a mean period of 53 (range 27-76) months. The patients were divided according to their initial response to systemic treatment with bromhexine. KCS parametres and p-IgG were measured repeatedly during the observation period. Patients responding to and continuously treated with bromhexine (2/3 of patients) improved significantly (P less than 0.05) in rose bengal score, but had increasing levels of p-IgG. Non-responders kept their low tear-production rate and had also increasing p-IgG levels. However, when subdivided according to p-IgG level, the group of patients with relatively low p-IgG improved in rose begal score, whereas the high p-IgG-group increased in rose bengal score. The rose bengal score appears to be a useful key parameter when evaluating disease level and progression.

Adult↗
PubMed1978

Bovine immunoglobulins and brucellosis. 3. Activity of IgG, IgG2 and IgM versus different commercial batches of Rose Bengal antigen.

The serological diagnosis of bovine Brucellosis by the Rose Bengal test has been disputed on account of the amount of active immunoglobulins and the results obtained in field trials. In order to try and explain these discrepancies, we have compared the activity of different commercial preparations of the Rose Bengal antigen for purified IgG1, IgG2 OR Igm dies. Two antigens which give the same titer for the Ig1 differ by a factor 8 in the titration of the IgM. Some antigens titer the IgG2, others, do not at all, or very little. Heifers in the 6th month of gestation were experimentally infected with Brucella abortus and half the animals showed "positive" from the 12th day with one antigen, whereas with another, they did not register until the 30th day. Heifers vaccinated at the age of 6-8 months with the B19 which still react to the Rose Bengal test 14 months later are 4 times more numerous with one of the antigens than with another.

Age Factors↗
PubMed1992

Detection of early xerophthalmia by impression cytology and rose Bengal staining--a comparative study.

Twenty two cases were studied for early detection of Xerophthalmia by impression cytology and Rose Bengal staining. Patients were below 6 years of age and admitted in the paediatric ward of Dhaka Medical College Hospital. Assessment were done by conjunctival impression cytology and staining with 1% Rose Bengal dye. Estimation of serum retinol level was done by HPLC to correlate conjunctival findings with biochemical status. The mean age of the subjects were 3.15 +/- 1.75. Rose Bengal staining was positive in 40% cases and impression cytology was in 60% cases. The mean serum retinol level of the diseased children were 10.5 +/- 3. ug/dl. The sensitivity of Rose Bengal staining was 53.3% and that of impression cytology was 80% in detecting vitamin A deficiency when compared with serum retinol level. The specificity of Rose Bengal staining was 40% and that of impression cytology was 100%. Sensitivity of impression cytology was 100% in patient with retinol level 10 ug/dl.

Child, Preschool↗
PubMed1986

A completely automatic method of processing 131I-labelled Rose Bengal dynamic liver studies.

A completely automatic method of measuring Rose Bengal uptake by the liver, expressed in terms of the half-time T1/2, is described. There is no requirement to construct time-activity curves with blood background activity correction as in the conventional regions-of-interest method. All the dixels in the image of a study are used in the actual data analysis. The method is therefore independent of operator influence. The intersection method is offered as an alternative to the manual method. The intersection method uses principal components analysis as a first step in the computation of the intersection of a theory space and a study space. A simple exponential function is used to generate the liver theory space. Sixty 131I Rose Bengal liver function studies were processed by the intersection method. The first forty minutes of patient data were used in the analysis. To validate the new method, computed T1/2 values were compared with those obtained by the manual method. A standard statistical test showed no significant difference between the two methods. Regression analysis gave a value for the coefficient of correlation of 0.89. The intersection method is currently in routine use for the automatic analysis of Rose Bengal liver studies and is faster than the manual method.

Autoanalysis↗
PubMedDate not supplied

Still confused about rose bengal?

We have surveyed the ophthalmic literature of the last five years in an attempt to evaluate the use and usefulness of Rose Bengal staining as an aid to differential diagnosis in dry eyes. Included both as a criterion and as an adjunct measure of disease progression, Rose Bengal scores of patients with different dry eye conditions overlap, sometimes to a considerable extent. A mechanistic link between staining with this dye and disease etiology is unlikely; however, Rose Bengal could be a surrogate marker of changes in ocular surface physiology. The question whether the extent and pattern of staining with Rose Bengal provide the clinician with information not available from other tests, and in particular from fluorescein staining of the ocular surface, has to be answered positively, though the nature of this information is not clearly understood. A more widespread recognition that Rose Bengal is not a vital dye is necessary in order not to bias the interpretation of experimental results.

Biomarkers↗
PubMed1993

Cellular stress by light and rose bengal in human lymphocytes.

Human lymphocyte cultures were supplemented with 10(-8)-10(-4) M Rose Bengal and irradiated with fluorescent light (Philips 40-W daylight-fluorescent lamp, 380-550 nm) and chromosomal aberrations and catalase and superoxide dismutase activities in the cells were determined. Chromosomal lesions and both enzymatic activities increased additively or synergistically in human lymphocytes after Rose Bengal supplementation and fluorescent light irradiation. Chromosomal lesions (expressed as chromosomal aberrations/cells) were (a) 0.12 +/- 0.03, (b) 0.18 +/- 0.12, (c) 1.58 +/- 0.11 and (d) 3.20 +/- 0.17 in the following conditions: (a) control; (b) after 3 h light irradiation, (c) supplemented with 10(-5) M Rose Bengal and (d) with dye treatment and light irradiation. Superoxide dismutase activity was: (a) 16.5 +/- 1.5; (b) 19.5 +/- 1.2; (c) 29.2 +/- 1.5 and (d) 35.4 +/- 2.1 U/mg protein and catalase activity was: (a) 10.3 +/- 0.5, (b) 12.8 +/- 0.7, (c) 22.4 +/- 0.5 and (d) 27.6 +/- 1.1 U/mg protein in the same experimental conditions. These findings suggest that Rose Bengal supplementation plus fluorescent light irradiation of human lymphocytes lead to the synthesis of superoxide dismutase and catalase in a manner similar to the heat-shock response. A threshold of chromosomal damage (about 2 chromosomal aberrations/cell) is apparently required to activate oxidative stress genes.

Adult↗
PubMed1983

Rose Bengal mediated inhibition of DNA polymerases: mechanism of inhibition of avian myeloblastosis virus reverse transcriptase under photooxidative conditions.

DNA polymerases from prokaryotic, eukaryotic, and oncornaviral sources are irreversibly inactivated upon exposure to Rose Bengal in the presence of light (photooxidation conditions). Inactivation of these enzymes under dark (nonoxidative) conditions is totally reversible [Srivastava, S. K., & Modak, M. J. (1982) Biochemistry 21, 4633-4639]. The primary effect of photooxidation on the enzyme-Rose Bengal complex was found to be the loss of template-primer binding ability within 5 min of exposure to light. The presence of template-primer but not the substrate deoxynucleotides consistently provided partial protection against Rose Bengal mediated photooxidative inactivation. Preformed enzyme-template-primer complexes were not dissociated by Rose Bengal under these conditions. However, prolonged exposure (25-30 min) of this complex to light in the presence of Rose Bengal led to complete inactivation of catalytic activity without affecting the ability of enzyme to bind to template-primer. These results clearly indicate that oxidative inactivation of avian myeloblastosis virus reverse transcriptase in the presence of Rose Bengal involves a domain within the enzyme that contains the template-primer binding site as well as an additional site which is required for the expression of both the polymerization and nuclease activities of the reverse transcriptase.

Avian Leukosis Virus↗
PubMed1989

Laser intensity and wavelength dependence of Rose-Bengal-photosensitized inhibition of red blood cell acetylcholinesterase.

The intensity and wavelength-dependence of Rose-Bengal-mediated photoinhibition of red blood cell acetylcholinesterase has been studied. Irradiation of dye-membrane suspensions with 308 nm laser excitation resulted in enzyme inhibition almost 50% greater than that obtained with 514 nm laser excitation. Sodium azide and argon purging greatly decreased the photosensitized enzyme inhibition at both wavelengths. Although Rose Bengal photosensitized enzyme inhibition more efficiently upon excitation into Sn (308 nm) than into S1 (514 nm), Stern-Volmer analysis of sodium azide quenching data gave similar quenching efficiencies at both wavelengths. Irradiation of dye-membrane suspensions with increasing intensities (Nd:YAG, 532 nm, 40 ps pulse duration) resulted in a decrease in enzyme inhibition. Saturation of the Rose Bengal fluorescence intensity and light transmission occurred with nearly the same intensity-dependence, suggesting that ground-state depletion occurs at the higher intensities. Our results demonstrate that excitation of a sensitizer into higher-lying excited singlet states can result in enhanced sensitizing efficiency. However, attempts to populate such states in Rose Bengal by sequential two-photon absorption using high intensities resulted only in ground-state depletion.

Cholinesterase Inhibitors↗