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Comparative studies on the tolerance to photoinduced cutaneous inflammatory reactions by psoralen and rose bengal.

The photochemotherapeutic value of topical 8-methoxypsoralen (8-MOP) plus UVA irradiation has been well recognized. The phototoxicity associated with psoralen plus UVA (PUVA) therapy is hallmarked by an increase in vascular permeability (iVP), the accumulation of polymorphonuclear leukocytes (aPMN) and erythema formation in situ. Rose bengal (RB) plus UVA-VIS light (320-700 nm) produces a similar acute inflammatory response, but without immediate or delayed erythema and perceptible edema. This study describes some of the parameters involved in inflammatory reactions evoked by PUVA and the results are compared with RB-induced phototoxic reactions. The rates of iVP and aPMN with a 3 h pulse were quantified using 125I-albumin and 51Cr-labelled PMNs respectively. The erythemal response was graded visually. 8-MOP cream was applied topically, while RB was injected intradermally in rabbit skin before UVA-VIS (9.4 J cm-2) irradiation. The data show that there is no significant difference in the rates of iVP, aPMN and erythema formation between normal skin sites and mast cell-depleted skin sites when challenged with 8-MOP plus light. These results suggest that in situ mast cells do not play a significant role in 8-MOP-photoinduced acute cutaneous inflammatory reactions, in contrast with RB-photoinduced reactions. The iVP and aPMN responses are minimal or absent in sites subjected to repeated exposure to 8-MOP plus light for three or more consecutive days, suggesting the establishment of a desensitized/unresponsive state. Moreover, 8-MOP-photo-desensitized sites do not produce iVP and aPMN of the same magnitude as the normal (naive) skin sites when challenged with RB plus light. Similarly, RB-photo-desensitized sites do not produce iVP and aPMN of the same magnitude as the native skin sites when challenged with 8-MOP plus light. The desensitization and cross-desensitization of skin sites to 8-MOP- or RB-photoinduced reactions suggest that there is either direct attack on the target cell(s), thereby removing the ability to express adhesion molecules, such as endothelial leukocyte adhesion molecule 1 (ELAM-1) or intercellular adhesion molecule 1 (ICAM-1), involved in the accumulation of inflammatory cells, or downregulation of the secretion/release of putative agent(s), such as interleukin 1 (IL-1) and tumor necrosis factor alpha (TNF-alpha), responsible for the initiation and progression of cutaneous inflammations.

Animals↗

Comparative study among complement fixation, serum agglutination and Rose Bengal Plate tests in the serodiagnosis of bovine brucellosis.

The complement fixation test (CFT) based on the 50 per cent hemolytic end point technique was applied to the diagnosis of bovine brucellosis in a comparative study involving the plate agglutination test (PAT), the tube agglutination test (TAT) and the Rose Bengal plate test (RBPT). Among 180 bovine sera that were examined the CFT was able to reveal significant titres in sera that gave low agglutinating titres. Some sera with agglutinating titres between 1 in 50 and 1 in 100 did not given a titre in the CFT. The results show that the CFT is useful for bovine brucellosis diagnosis and may be used after sera screening with PAT and RBPT.

Agglutination Tests↗

Comparative study among complement fixation, serum agglutination and Rose Bengal plate tests in the serodiagnosis of bovine brucellosis.

The complement fixation test (CFT) based on the 50 per cent hemolytic end point technique was applied to the diagnosis of bovine brucellosis in a comparative study involving the plate agglutination test (PAT), the tube agglutination test (TAT) and the Rose Bengal plate test (RBPT). Among 180 bovine sera that were examined the CFT was able to reveal significant titres in sera that gave low agglutinating titres. Some sera with agglutinating titres between 1 in 50 and 1 in 100 did not give a titre in the CFT. The results show that the CFT is useful for bovine brucellosis diagnosis and may be used after sera screening with PAT and RBPT.

Agglutination Tests↗

Inactivation of NADP(+)-dependent isocitrate dehydrogenase by singlet oxygen derived from photoactivated rose bengal.

Recently, we demonstrated that the control of cytosolic and mitochondrial redox balance and the cellular defense against oxidative damage is one of the primary functions of NADP(+)-dependent isocitrate dehydrogenase (ICDH) by supplying NADPH for antioxidant systems. When exposed to a singlet oxygen-producing system composed of rose bengal (RB) and visible light, ICDH was susceptible to oxidative modification and damage as indicated by the loss of activity and by the formation of carbonyl groups. The structural alterations of modified enzyme were indicated by the increase in susceptibility to proteases and the change in intrinsic fluorescence spectra. Upon exposure to photoactivated RB, a significant decrease in both cytosolic and mitochondrial ICDH activities was observed in HL-60 cells. The singlet oxygen-mediated damage to ICDH may result in the perturbation of cellular antioxidant defense mechanisms and subsequently lead to a pro-oxidant condition. When we examined the antioxidant role of cytosolic ICDH against singlet oxygen-induced damage with HL-60 cells transfected with the cDNA for mouse cytosolic ICDH in sense and antisense orientations, a clear inverse relationship was observed between the amount of cytosolic ICDH expressed in target cells and their susceptibility to singlet oxygen-mediated oxidative damage.

Animals↗

Photosensitizers neutral red (type I) and rose bengal (type II) cause light-dependent toxicity in Chlamydomonas reinhardtii and induce the Gpxh gene via increased singlet oxygen formation.

The connection between the mode of toxic action and the genetic response caused by the type I photosensitizer and photosynthesis inhibitor neutral red (NR) and the type II photosensitizer rose bengal (RB) was investigated in the green alga Chlamydomonas reinhardtii. For both photosensitizers, a light intensity-dependent increase in toxicity and expression of the glutathione peroxidase homologous gene (Gpxh) was found. The toxicity of RB was reduced by the singlet oxygen (1O2) quenchers 1,4-diazabicyclo[2.2.2]octane and L-histidine, and the RB-induced Gpxh expression was stimulated in deuterium oxide-supplemented growth medium. These observations clearly indicate the involvement of 1O2 in both toxicity and the genetic response caused by RB. NR up-regulated the expression of typical oxidative and general stress response genes, probably by a type I mechanism, and also strongly induced the Gpxh expression. The stimulating effect of deuterium oxide in the growth medium suggested the involvement of 1O2 also in the NR-induced response. Indeed, an increased 1O2 formation was detected with EPR-spin trapping in NR-treated spinach thylakoids. However, none of the 102 quenchers could reduce the light-dependent toxicity of NR in C. reinhardtii, indicating that NR has a different mode of toxic action than RB.

Animals↗

Systematic study of parameters influencing the action of Rose Bengal with visible light on bacterial cells: comparison between the biological effect and singlet-oxygen production.

As part of a project to study different methods for the disinfection of effluent water, the inactivation of different microorganisms (Escherichia coli, Deinococcus radiodurans and spores of Bacillus subtilis) using a combination of a photosensitizer (Rose Bengal) with simulated sunlight and oxygen was determined under various environmental conditions (temperature, pH index). In parallel, the singlet-oxygen (1O2) production was also measured under the same conditions. Whereas the vegetative cells could be inactivated much more efficiently at increased temperature and altered index of pH, the production of 1O2 remained essentially the same under these alterations. Additionally, the relations among the sensitivities of different cell types to be killed by our photodynamic treatments (PDT) were opposite to those found after exposure to ionizing radiation. The results of photodynamic experiments do not reflect the cells' capacity to repair DNA strand breaks. Spores of B. subtilis, as a nonvegetative system, could not be inactivated by illuminations up to 100 J cm-2. Together, these findings indicate that DNA is not the primary target, the inactivation of which leads to the killing of our test organisms. Instead, the cellular envelope appears to be the component being assaulted by our PDT.

Bacteria↗

Suitability criteria for compartmental analysis of the plasma clearance curve of exogenous cholephils. 131I Rose Bengal fulfils these criteria.

Hepatic clearance of exogenous cholephils involves at least three transfers : uptake by the liver, reflux from hepatocytes into the plasma, and intracellular transport followed by active secretion in bile. Compartmental analysis may be applied to the plasma disappearance curve in order to calculate the fractional transfer rates. The 131I Rose Bengal (131I RB) fills the requirements of such an analysis, contrary to bromsulphalein and indocyanine green, which have been used until now. 131I RB does not undergo enterophepatic cycle and is not metabolized by the liver. It behaves, in the preparation used, like a homogeneous substance, remains stable in the serum as long as 25 hours after its injection and may be bound completely with human albumin in vitro prior to administration : under these conditions, the material which is cleared by the liver is homogeneous. The use of tracer instead of loading doses, avoid any toxic effects on transport systems and results in plasma and liver concentrations which are negligeable in comparison with the Km of membrane transport, and are compatible with first order kinetics.

Chromatography↗

Photoelectrocatalytic degradation of Rose Bengal.

An innovative photoelectrode, TiO2/Ti mesh electrode, was prepared by galvanostaticanodisation. The morphology and the crystalline texture of the TiO2 film on mesh electrode were examined by scanning electronic microscopy and Raman spectroscopy respectively. The examination results indicated that the structure and properties of the film depended on anodisation rate, and the anatase was the dominant component under the controlled experimental conditions. Degradation of Rose Bengal (RB) in photocatalytic (PC) and photoelectrocatalytic (PEC) reaction was investigated, the results demonstrated that electric biasing could improve the efficiency of photocatalytic reaction. The measurement results of TOC in photoelectrocatalytic degradation showed that the mineralisation of RB was complete relatively. The comparison between the degradation efficiency of RB in PEC process and that in aqueous TiO2 dispersion was conducted. The results showed that the apparent first-order rate constant of RB degradation in PEC process was larger than that in aqueous dispersion with 0.1%-0.3% TiO2 powder, but was smaller than that in aqueous dispersion with 1.0% TiO2.

Catalysis↗

Antiretroviral activities of hypericin and rose bengal: photodynamic effects on Friend leukemia virus infection of mice.

The ability of hypericin to protect mice from splenomegaly resulting from infection with Friend leukemia virus (FLV) was re-examined in light of recent evidence showing that light is absolutely required for this drug's antiviral activity. FLV-induced splenomegaly was not prevented or ameliorated in mice injected with 100 micrograms hypericin, either mixed with the FLV inoculum or administered 1 day p.i., either under normal laboratory light or in the dark. These results contradict previous findings. Both hypericin and rose bengal, however, inactivated the FLV inoculum at low doses (< 11 micrograms), provided that the mixture was illuminated for 1 h under a normal fluorescent desk lamp. This procedure protected mice completely from FLV-induced splenomegaly, and provided a possible explanation for the discrepancy between our results and those reported previously. We conclude that for FLV, as for other enveloped viruses studied previously, illumination of hypericin with the virus is absolutely required for hypericin's antiviral (virucidal) effects, thus limiting its potential usefulness as an antiretroviral agent.

Animals↗

The inhibition of glutamate dehydrogenase by L-serine O-sulphate and related compounds and by photo-oxidation in the presence of Rose Bengal.

1. Glutamate dehydrogenase was inhibited by l-serine O-sulphate, beta-chloro-l-alanine, O-phospho-l-serine and beta-chloro-l-alanine methyl ester. With the exception of beta-chloro-l-alanine methyl ester which was an irreversible inhibitor, it was possible to reverse the inhibitory effects by dialysis. 2. Both NAD(+) and glutamate afford some protection against the inhibition due to the methyl ester. No change in the normal stimulatory effect exhibited by ADP was observed in the presence of beta-chloro-l-alanine methyl ester but the effect due to GTP was modified. 3. Irradiation of glutamate dehydrogenase in the presence of Rose Bengal produced rapid inactivation. Amino acid analysis of the inactivated enzyme showed that eight histidine residues had been destroyed in the process.

Adenosine Diphosphate↗

Firefly luciferin-activated rose bengal: in vitro photodynamic therapy by intracellular chemiluminescence in transgenic NIH 3T3 cells.

Photodynamic therapy (PDT) of cancer (1, 2) is a well-established treatment modality that uses light excitation of a photosensitive substance to produce oxygen-related cytotoxic intermediates, such as singlet oxygen or free radicals (3, 4). Although PDT is advantageous over other forms of cancer treatments because of its limited side effects, its main disadvantage is the poor accessibility of light to more deeply lying malignancies. External light sources such as lasers or lamps can be applied either noninvasively to reach tumors that lie well within the penetration depth of the light or in a minimally invasive fashion (interstitial treatments) in which optical fibers are placed intratumorally through needles. Even with the second approach, light distribution over the tumor is not homogeneous and nonidentified metastatic disease is left untreated. CL, the chemical production of light, is exemplified by firefly light emission mediated by the enzymatic (luciferase + ATP) oxidation of D-luciferin to oxyluciferin (5). This mobile light source is a targetable alternative to external sources of illumination. Here we show the in vitro photodynamic effect of rose bengal activated by intracellular generation of light, in luciferase-transfected NIH 3T3 murine fibroblasts.

3T3 Cells↗

Inactivation of leucine aminotransferase with diethylpyrocarbonate and rose bengal: evidence for an active site histidine residue.

Modification of leucine aminotransferase by diethylpyrocarbonate or rose bengal-sensitized photo-oxidation caused rapid inactivation of the enzyme. The inactivation of leucine aminotransferase depended on the concentration of the reagent, the time of incubation and exhibited pseudo-first order kinetics. Rose bengal-sensitized photo-oxidation was maximum at pH 6.5 and 9. Substrates leucine and alpha-ketoglutarate protected the enzyme against inactivation by these reagents, thus suggesting participation of histidine residue at the substrate binding site.

Binding Sites↗

Influence of dynamic contact of hard contact lens materials on corneal epithelial cells examined by rose bengal staining.

PURPOSE: To establish a method for evaluation of less irritating contact lens materials by dynamic contact with cornea, we examined epithelial and endothelial cell injury to the porcine cornea caused by rotatory rubbing with four kinds of hard contact lenses (HCL). METHODS: The HCLs used were (1) polymethylmethacrylate (PMMA) HCL, (2) gas-permeable HCL composed of a graft co-polymer of dextran derivative and methylmethacrylate (MMA) (Suncon Mild II(TM), 12 Dk), (3) gas-permeable HCL composed of a graft co-polymer of dextran derivative, a monomer containing silicone, a monomer containing fluorine and MMA (New Dx HCL-136, 32 Dk), and (4) gas-permeable HCL composed of a monomer containing silicone, a monomer containing fluorine and MMA(RGPL-A, 216 Dk). Using a specially designed apparatus, we produced a standardized injury to the epithelium or endothelium of porcine corneas by holding the HCL against the corneal surface while rotating the lens rapidly. RESULTS: After rotatory rubbing of the HCL on the epithelium, the degree of rose bengal staining of the epithelial cells, indicating degeneration of the cells and mucin detachment, were significantly different among these HCLs (New Dx HCL-136 < Suncon Mild II(TM), PPMA < RGPL-A), and the cell injury rates of the endothelium were also significantly different among them (Suncon Mild II(TM) < New Dx HCL-136, PMMA < RGPL-A). The water-wettability of HCLs was not directly correlated with cell injury rates on epithelial and endothelial cells. CONCLUSIONS: Both the New Dx HCL-136 and Suncon Mild II(TM), which have a common composition of graft co-polymer of dextran derivative, are less irritating to the epithelium and to the endothelium. Also a very few patients complaints regarding Suncon Mild II(TM) wearing in individuals with dry eyes have been reported. Therefore, we would expect that the New Dx HCL-136 should be satisfactory for wear by individuals with dry eyes.

Animals↗