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The I-131 rose bengal excretion test is not dead.

One hundred and thirty I-131 Rose Bengal Excretion Studies (RBI) were performed on 84 patients over nine years. In 90% (56/60) of cases with biliary atresia, the 72-hour RBI was less than or equal to 7%. In only 12.5% (3/24) of cases with neonatal hepatitis was the 72-hour RBI less than or equal to 7%. The accuracy of the test was 91% with a specificity of 88%. Thirty patients later were studied following a Kasai procedure. The RBI test reliably predicted the patency of the anastomosis. The authors conclude that the 72-hour RBI is a reliable test in the diagnosis of biliary atresia and in the documentation of biliary patency following surgery, provided adequate care is taken in stool collection and measurement.

Bile Ducts↗

[Hepatic scintigraphy using 123I-rose bengal].

The paper is devoted to the experience in diagnostic investigations using rose bengal, labeled with half-life 123I (123I-RB). The drug was synthesized in the B. P. Konstantinov Leningrad Institute of Nuclear Physics, USSR Academy of Sciences. Analysis of the results of investigations of 201 patients with diseases of the liver and biliary tracts revealed the advantages of 123I-RB over standard RPs of similar diagnostic purpose. Owing to low radiation exposure one could use higher activity (up to 120 MBq and more) of the drug. It made it possible to combine a study of the functional parameters of the hepatobiliary system and obtaining scintigraphic data on the structure of the liver and biliary tracts. 123I-RB is the best radiodiagnostic agent for a study of function and topography of the liver in patients with an increased blood level of bilirubin.

Adult↗

Topical rose bengal: pre-clinical evaluation of pharmacokinetics and safety.

BACKGROUND AND OBJECTIVES: Rose bengal (RB) is a potent photosensitizer that has largely been overlooked as a potential photodynamic therapy (PDT) agent. In this study, the feasibility of topical delivery of RB to the epidermis has been evaluated. STUDY DESIGN/MATERIALS AND METHODS: Topical formulations of RB were assessed on murine and rabbit skin for pharmacokinetic properties, cutaneous toxicity, and photosensitization. RESULTS: Hydrophilic formulations (<or=1% RB) exhibited rapid, selective, uniform delivery to the epidermis, with no significant acute cutaneous toxicity in normal skin. Illumination (532 nm) elicited no acute phototoxicity for light intensities <or=100 mW/cm(2) at a light dose of 100 J/cm(2); use of higher intensities resulted in superficial thermal damage. Repeat treatment of rabbit skin (weekly for four weeks) elicited minor phototoxicity only at the highest concentration (1% RB). CONCLUSIONS: These results indicate that RB is safe for PDT treatment of skin disorders, exhibiting negligible effects in normal skin.

Administration, Cutaneous↗

[Diagnostic value of the rose bengal plate test in the diagnosis of bovine brucellosis].

The value of rose bengal plate test (RBPT) in diagnosing brucellosis in cattle was determined by statistical comparison of its results with the results of the tests used in Poland, i.e. SAT, CFT, AGT and MET. RBPT was made in 2 variants. In routine, highly specific test, equal parts of antigen and serum--0.03 cm3, were used whereas in the experimental one the sensitivity was increased using half the amount of antigen--0.015 cm3 (RBPT0.015). Two batches of cattle serum were examined. In group I 249 cattle serums from the herds infected with brucellosis were examined. In group 2 there were 1269 cattle serums from the herds free of brucellosis, positive in SAT but negative in CFT. The reactions in SAT were considered as not specific if the reaction in the additional examination in CFT, AGT and MET was negative. On the basis of the results in group I, mainly the sensitivity of RBPT was determined compared with the total evaluation of the results of SAT/CFT. In RBPT0.015 the consistency of the results was 99.4% but in RBPT0.03 only 87.9%. Detectability of reactions, i.e. the percentage of positive results in infected herds was calculated. The results were as follows: AGT--89.6%, RBPT0.015--74.3%, SAT/CFT--66.3%, CFT--65.9%, RBPT0.03--59.8%, SAT--55.4%. In the group 2 mainly specificity of RBPT in relation to CFT, AGT and MET was determined. In RBPT0.03 it achieved 97.1% whereas in RBPT0.015--83.1%. The application of RBPT0.03 in the group 2 eliminated 95.8% of the not specific reactions in SAT and that of RBPT0.015--77.9%, respectively. The author suggests to use RBPT0.03 as a screening method instead of SAT and CFT to diagnose cattle brucellosis in the areas free from the disease. On the other hand, RBPT0.015, as more sensitive test is suggested to be used in the herds suspected of brucellosis to identify quickly infected animals.

Agglutination Tests↗

Dichloran-rose bengal medium for enumeration and isolation of molds from foods.

Overgrowth by spreading molds such as Rhizopus and Mucor species is a problem with fungal enumeration media used for foods. Thirty-one antifungal compounds were surveyed for their ability to selectively inhibit such fungi while allowing growth of mycotoxigenic molds and other species of significance in food spoilage. Dichloran (2,6 dichloro-4-nitroaniline) restricted growth of Rhizopus stolonifer while allowing satisfactory growth of the other test molds. Three Rhizopus and Mucor species were encountered that were not inhibited by dichloran; these were controlled by the addition of rose bengal. The optimal medium, designated DRBC, contained 2 micrograms of dichloran and 25 micrograms of rose bengal per ml. DRBC, in pure culture tests and with food samples, restricted the colony size of spreading molds and recovered a wider range of species in higher numbers than other enumeration media.

Aniline Compounds↗

Evaluation of rose bengal staining test and rapid dark-adaptation test for the field assessment of vitamin A status of preschool children in Southern Brazil.

The objective of this study was to evaluate two of several methods presently available for assessing vitamin A status of marginally malnourished preschool children of socioeconomically deprived families in Southern Brazil. The rose bengal staining test and rapid dark-adaptation test were evaluated by comparing their results with conventional methods that require estimation of dietary intake of vitamin A equivalents and determination of plasma levels of retinol before and after an oral supplementation of 200 000 IU vitamin A. Results indicate that neither test evaluated is useful by itself. There was no significant correlation between the results of the rose bengal staining test and the plasma levels of retinol. Also, no significant correlation was observed between the rapid dark-adaptation time and the vitamin A status of the preschool children as determined by plasma-retinol levels before and after vitamin A supplementation.

Brazil↗

Effects of phenobarbital and rose bengal on the ATPases of plasma membranes of rat and rabbit liver.

The effects of drugs which change the bile-salt-independent fraction of bile flow on Na(+)K(a+) and Mg(2+) activated ATPases were studied in membrane fractions rich in bile canaliculi. The administration of phenobarbital caused no induction of these enzymes which could explain the increased bile flow observed in the rat. Rose bengal, in addition to its strong photoxidative inhibition of both ATPases, inhibits the Na(+)K(+) ATPase of rat and rabbit bile canaliculi in the absence of light. A closely related substance, uranine, inhibits neither bile flow nor Na(+)K(+)ATPase. Inhibition of this enzyme by rose bengal may therefore be responsible for the observed effects of this dye on bile flow independent of bile salts.

Adenosine Triphosphatases↗

Renal uptake of 131I-rose bengal in biliary atresia. A case report.

Renal uptake of radio-iodinated rose bengal was studied in a 5-month-old infant, who was later found, at operation, to have biliary atresia. Prominent renal uptake, which could easily be mistaken for radioactivity in the bowel, was noted. While this pitfall in diagnosis has previously been described, it is thought worth while to stress this possible cause of misinterpretation.

Bile Ducts↗

131I Rose Bengal scanning and clearance ratios in the investigation of jaundiced patients.

It may be difficult to distinguish between obstructive and non-obstructive jaundice despite the use of clinical, biochemical and radiographic tests. Endoscopic cannulation tests promise to be the most accurate means of separating the two groups, but they are not yet widely available, and are not entirely safe. In contrast, the 131I-Rose Bengal test is simple, safe and comparatively widely available. Its accuracy in 70 jaundiced patients is reviewed to determine whether it still has a place in the investigation of jaundiced patients. Blood clearance traces, intestinal radioactivity traces and serial abdominal scans were done during a period of 3 h. The blood clearance ratios were not helpful discriminators, and the most helpful information was the timing of intestinal radioactivity. Its rapid appearance excluded mechanical obstruction. A delay of up to 3 h in its appearance was characteristic of cholestasis. A delay of more than 3 h was found in all cases of mechanical obstruction, but also in ten cases of severe cholestasis which could not be distinguished from those with mechanical obstruction. A follow-up examination within another ten days was of diagnostic value in two of these cases and the overall accuracy of the test was 86%. The potential value of prolonging the test to 24 h is discussed. It is concluded that the 131I-Rose Bengal test is a helpful diagnostic discriminator in the great majority of patients with jaundice. It is offered as a screening procedure for specialised techniques.

Biliary Tract Diseases↗

Evidence against singlet oxygen formation by sonolysis of aqueous oxygen-saturated solutions of Hematoporphyrin and rose bengal. The mechanism of sonodynamic therapy.

The possible role of singlet oxygen in the mechanism of sonodynamic therapy, the synergistic effect of ultrasound and certain sonosensitizers, was investigated. We used 4,4'-bis(1-p-carboxyphenyl-3- methyl-5-hydroxyl)-pyrazole (DRD 156), a sensitive new reagent which reacts specifically with singlet oxygen (1O2) but not with OH radicals, superoxide anion radicals or H2O2, to produce an EPR detectable signal. Sonolysis (48 kHz) of 90% D2O oxygen-saturated PBS solutions of Hematoporphyrin or Rose Bengal did not lead to the formation of detectable EPR signals of the semiquinone radical of DRD156. In contrast, the EPR signal of the semiquinone radical of DRD156 was observed during photoirradiation of Hematoporphyrin at 505 nm or of Rose Bengal at 544 nm. These results are inconsistent with a major role for singlet oxygen formation in the sonolysis of aqueous solutions of these compounds. An alternative mechanism for sonodynamic therapy involving peroxyl and alkoxyl radicals is discussed.

Electron Spin Resonance Spectroscopy↗

Partition of rose bengal anion from aqueous medium into a lipophilic environment in the cell envelope of Salmonella typhimurium: implications for cell-type targeting in photodynamic therapy.

Photodynamic therapy employs photosensitizers for the selective destruction of tumor tissue while sparing the surrounding healthy tissue. Photosensitization may also be applied to the selective eradication of microorganisms. Photosensitized inactivation requires that the sensitizer bind to the target and therefore the factors that determine photosensitizer binding are critical to photosensitization selectivity. This paper reports the determination of some features of the binding site of the potent photosensitizer, Rose Bengal, in Salmonella bacteria and describes some of the factors that affect this binding. The shift in the wavelength of maximum fluorescence and experiments with the fluorescence quencher TNBS indicate that Rose Bengal is located in a non-aqueous compartment such as the outer membrane. The dye does not seem to significantly accumulate inside the cell, but rather to accumulate in the outer membrane. Time-dependent changes in sensitizer localization in two strains of Salmonella typhimurium that differ in cell wall formation, LT-2 and TA1975, correspond to their differences in susceptibility to photosensitized killing. Therefore these results provide clues to the factors that determine photosensitization selectivity. Understanding this phenomenon is essential for the efficient design of selective photosensitizers and for optimizing antitumor and antiviral photodynamic therapy.

Binding Sites↗

The serological diagnosis of bovine brucellosis: an evaluation of the complement fixation, serum agglutination and rose bengal tests.

In a collaborative investigation in which 4 laboratories took part, the Rose Bengal Test (RBT), Serum Agglutination Test (SAT) and 4 different Complement Fixation (CFT) techniques were evaluated in selected cattle for the diagnosis of bovine brucellosis, by comparing the results they gave with the bacteriological examination of a selection of lymph nodes taken from the same animals at slaughter. The RBT correctly classified all but 1 of 79 culture-positive cattle, but was more often positive in culture-negative animals than the other tests. The RBT may be most useful as a screen test. 11% of the culture-positive cattle had SAT titres below 100 iu and almost 4% of them had less than 30 iu. However, the SAT was more effective in cattle vaccinated with the 45/20 vaccine. The CFT, in 1 of 4 techniques used, identified all of the culture-positive cattle at a serum dilution of 1/4 or above and was considered to be far superior to the SAT as a diagnostic test, except perhaps in cattle vaccinated with the 45/20 vaccine.

Agglutination Tests↗

Elicitation of spreading depression by rose bengal photodynamic action.

Spreading depression refers to a slowly propagating depression of the ordinary electrical activity of the nervous tissue. It can be elicited by different types of physical or chemical non-specific stimuli. Various evidences suggest that transient alterations of cell membranes are involved. For this reason, and considering the action of free radicals on cell membranes, the elicitation of the reaction by dye photoactivation has been investigated. Isolated chick retina superfused in the dark with Ringer solution was able to regularly exhibit spreading depression when submitted to 1 microM rose bengal pulse of 5 min in duration, followed by 2.1 x 10(4) to 4.2 x 10(4) Jm-2 light pulse. The phenomenon was monitored either by visual inspection of the light-scattering milky wave that accompanies the reaction or by recording its characteristic slow voltage variation. The reaction was not triggered if the retina, superfused with the dye, was (a) maintained in the dark; (b) illuminated with red light (3.75 x 10(2) to 2.25 x 10(4) Jm-2), or (c) stimulated by white light but superfused with nitrogen-saturated solutions. It is concluded that, under the present conditions, the elicitation of spreading depression is contingent on the photoactivation of rose bengal in the presence of oxygen.

Animals↗

Photodynamic action of rose bengal on isolated rat pancreatic acini: stimulation of amylase release.

The halogenated fluorescein derivative, rose bengal, upon photon activation, elicits amylase secretion from isolated, perifused pancreatic acini. This effect is due to production of highly reactive singlet delta oxygen which can permeabilize the cell membrane and may also react chemically with secretagogue receptors, or other functional components of the membrane such as the G-proteins. The profile of photodynamically induced amylase secretion is anion-dependent: it becomes biphasic when the chloride ion is substituted by the glutamate ion, an effect attributed to the action of glutamate on the ionic transport systems of the zymogen granule membrane.

Amylases↗

Rose Bengal adsorbed on microgranular cellulose: evidence on fluorescent dimers.

Rose Bengal adsorbed on microgranular cellulose was studied in the solid phase by total and diffuse reflectance and steady-state emission spectroscopy. A simple monomer-dimer equilibrium fitted reflectance data up to dye loadings of 4 x 10(-7) mol (g cellulose)(-1) and allowed calculation of monomer and dimer spectra. Further increase of dye loading resulted in the formation of higher aggregates. Observed emission and excitation spectra and quantum yields were corrected for reabsorption and reemission of luminescence, using a previously developed model, within the assumption that only monomers are luminescent [M. G. Lagorio, L. E. Dicelio, M. I. Litter and E. San Roman, J. Chem. Soc., Faraday Trans., 1998, 94, 419]. An apparent increase of fluorescence quantum yield with dye loading was found, which was attributed to the occurrence of dimer fluorescence. Extension of the model to two luminescent species (i.e. monomer and dimer) yielded constant fluorescence quantum yields for the monomer, phiM= 0.120 +/- 0.004, and for the dimer, phiD= 0.070 +/- 0.006. The monomer quantum yield is close to the value found for the same dye in basic ethanol. The presence of fluorescent dimers and calculated quantum yields are supported by analysis of the excitation spectra and other experimental evidence. The possible occurrence of non-radiative energy transfer and the effect of surface charge on the properties of the dimer are analyzed.

Journal Article↗

131I rose bengal: its use in the evaluation of infantile jaundice.

One-hundred ten 131I-rose bengal studies (RBI) were performed in infants suspected of having biliary atresia. Fecal RBI excretion of less than 10% was observed in 72 of 73 cases of extrahepatic biliary atresia, but also in 10 of 37 cases of intrahepatic cholestasis of various origins. One-hundred twenty-two RBI tests were performed in children operated on for extrahepatic biliary atresia and 71 tests were performed between postsurgical weeks 3 and 8, and 51 tests were done later. Prognostically, early tests show that fecal RBI excretion of more than 15% was observed in 2 of 34 cases who were later completely jaundice-free and in only 1 of 37 cases where no bile flow restoration occurred.

Bile Ducts↗

Rose bengal as a tool in studying the ligand binding of phosphorylase b.

The interaction of rose bengal (RB) with rabbit skeletal muscle phosphorylase b (1,4-alpha-D glucan: orthophosphate alpha-glucosyl-transferase, E.C. 2.4.1.1.) was studied by kinetic and absorption photometric methods. RB inhibited the phosphorylase b activity. Inhibition was strictly competitive with respect to substrate G-1-P and activator AMP with inhibition constants 2 x 10(-6) M and 2.2. x 10(-7) M, respectively. The association of the dye with the enzyme elicited a red shift in the spectrum of RB indicating an apolar binding site. According to difference absorption measurements, the enzyme binds two dye molecules per dimer in the presence and absence of both G-1-P and AMP. Binding constants determined from photometric titrations are consistent with those obtained from kinetic measurements. The present findings allow to carry out detailed kinetic investigations on the activator AMP and substrate G-1-P binding of phosphorylase b.

Adenosine Monophosphate↗