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Rose Bengal located within liposome do not affect the activity of inside-out oriented Na,K-ATPase.

DPPC:DPPE-proteoliposomes (in which the enzyme is inside-out oriented) and DLOPC:DLOPE-proteoliposomes (in which the enzyme is only 40% inside-out oriented) is an excellent model for studying the selective effect of the reactive oxygen species, produced by the photo-activation of Rose Bengal. Both proteoliposomes used, when submitted to photo-irradiation with laser using 1200 mJ/cm2 energy dose, in the absence of the Rose Bengal, did not shown any effect in the ATPase activity and in the integrity of its systems. Also, no effect was observed using 50 microM of Rose Bengal encapsulated in the interior of the DPPC:DPPE-proteoliposome system. But, when we use 50 microM of Rose Bengal, present only in the extravesicular environment, and photo-irradiation with a laser dose of 200 mJ/cm2, it results in the loss of 40-50% of the ATPase activity, with damage of the DPPC:DPPE-proteoliposome integrity. Using a dose of 400 mJ/cm2 the ATPase activity was totality lost. Consequently, these effects could be correlated with direct damage in the peptide structure. The photo-irradiation of the system constituted by DLOPC:DLOPE-proteoliposome in the presence of Rose Bengal, encapsulated only in the interior compartment or in the extra-liposomal environments, revealed a gradual decrease of the ATPase activity, maintaining it at 30% after a dose of 1200 mJ/cm2 and losing total ATPase activity at 800 mJ/cm2, respectively, with the loss of integrity of this vesicular system in both conditions studied. The generated singlet oxygen could attack the double linkage present in the fatty acid structure of the lipid instead of the amino acid in the protein structure and, in a second step, result in an indirect inactivation of the enzyme activity. In summary, these results indicated that singlet oxygen species produced by photo-oxidation of Rose Bengal using laser light could act in protein and lipid structure depending on its proportion or distribution.

Liposomes↗

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↗

Rose bengal dye concentration and volume delivered via dye-impregnated paper strips.

PURPOSE: The technique of administration and the resulting volume and concentration of the dye delivered to the eye via dye-impregnated paper strips are not standardized or known. The purpose of this study was to use a standard technique for preparation of rose bengal dye from commercially available paper strips to determine the variability in concentration based upon soak time, and to determine the volume delivered by such a technique to a model eye. METHODS: Rose bengal filter paper strips were wetted for either 15, 30, or 45 s, and the resulting concentration of dye eluted was determined by spectrophotometry. With the same preparation technique and a 15-s soak time, the strip was applied to wetted filter paper, as a model for the moist surface of the eye. The volume of solution delivered was calculated by weight. RESULTS: Concentrations of rose bengal for the various soak times were: 15 s: 7.64 (+/- 1.11) by 10(-6) g/ml; 30 s: 9.70 (+/- 1.79) by 10(-6) g/ml; and 45 s: 10.27 (+/- 0.66) by 10(-6) g/ml. The volume delivered to the eye model with this technique was 17.43 microliters (+/- 3.09). CONCLUSIONS: The concentration of rose bengal dye delivered to the ocular surface by means of a wetted strip is relatively low and soak time/technique-dependent. This suggests a possibility that results in clinical studies with rose bengal strip application may be different than if controlled volume applications of commercially available 1% liquid rose bengal dye are used.

Cornea↗

Effects of rose bengal on bile secretion in the rabbit: inhibition of a bile salt-independent fraction.

The authors have studied the influence of rose bengal, and, by comparison, of uranin, on choleresis in the rabbit. Uranin induced an increase in bile flow and a decrease in bile salt concentration in bile. These results are consistent with an osmotic mechanism, as proposed by Sperber (1959). By contrast, rose bengal induced a marked decrease in bile flow and an increase in bile salt concentration. In rose bengal-treated animals (30 mumoles per kilogram body weight intravenously), bile flow was 65% lower and bile salt concentration 95% higher than in control animals studied during the same periods. The relationship between bile flow and bile salt excretion is consistent in the rabbit, with the existence of a bile salt-independent secretion of 60 mg per minute per kilogram body weight, that is, 60% of bile flow. In rose bengal-treated animals, this fraction was only 15 mg per minute per kilogram body weight. It is concluded that rose bengal inhibits the bile salt-independent fraction. Rose bengal, as other phtalein dyes, is excreted by the liver cells. Its effect on the bile salt-independent fraction suggests that this fraction is secreted by the hepatocytes.

Animals↗

Rapid electrophysiological changes leading to arrhythmias in the aerobic rat heart. Photosensitization studies with rose bengal-derived reactive oxygen intermediates.

The objective of this study was to determine whether reactive oxygen intermediates (e.g., singlet oxygen and the superoxide radical) can rapidly induce electrophysiological disturbances leading to the genesis of arrhythmias, even in the absence of ischemia and reperfusion. Rat hearts (n = 6 per group) were perfused aerobically at 37 degrees C for 10 minutes without rose bengal and for 5 minutes with rose bengal (250 nmol/l), during which time no changes in coronary flow or heart rate were observed. Hearts were then uniformly illuminated for 20 minutes with green light (530-590 nm) from 200 fiber optic cables. With light and without rose bengal, or vice versa, all hearts remained stable. However, in the illuminated rose bengal group, electrophysiological changes (inversion of the terminal portion of the T wave and an increase in Q-T interval) were observed within 11.8 +/- 2.1 seconds (i.e., less than 60 beats). All hearts exhibited ventricular premature beats (within 2.2 +/- 0.7 minutes) and ventricular tachycardia (within 2.8 +/- 0.7 minutes) before the occurrence of complete atrioventricular block (within 5.5 +/- 0.9 minutes). During the illumination period, coronary flow progressively fell in the rose bengal-perfused hearts from 11.6 +/- 0.5 ml/min to 2.0 +/- 0.4 ml/min (p less than 0.05 when compared with any control group). When a similar progressive reduction in coronary flow was mimicked (with or without rose bengal), no arrhythmias occurred.(ABSTRACT TRUNCATED AT 250 WORDS)

Aerobiosis↗

[Combined use of rose Bengal and indirect immunofluorescence in the diagnosis of brucellosis].

In the present prospective study we have evaluated the sensitivity, specificity and predictive value of the Bengal rose and indirect immunofluorescence (IIF) in 122 patients with a bacteriological diagnosis of brucellosis. The sensitivity of the Bengal rose was 95.79% and its specificity 98.43%. IIF had a sensitivity of 68.80% and a specificity of 100% when the polyvalent anti-IgS conjugate was used. The combined parallel use of Bengal rose and IIF-IgS resulted in a sensitivity of 89.17% and a specificity of 100%. There was a good correlation between Bengal rose and serum agglutination test (r = 0.72); on the contrary, there were not good correlations between the fluorescent conjugates and Bengal rose, agglutination and Coombs test. In conclusion, the combined use of Bengal rose as a screening test and IIF as a confirmation study is a good diagnostic strategy for human brucellosis.

Agglutination Tests↗

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↗

Construction of an integrated map of rose with AFLP, SSR, PK, RGA, RFLP, SCAR and morphological markers.

A high-density genetic map with a number of anchor markers has been created to be used as a tool to dissect genetic variation in rose. Linkage maps for the diploid 94/1 population consisting of 88 individuals were constructed using a total of 520 molecular markers including AFLP, SSR, PK, RGA, RFLP, SCAR and morphological markers. Seven linkage groups, putatively corresponding to the seven haploid rose chromosomes, were identified for each parent, spanning 487 cM and 490 cM, respectively. The average length of 70 cM may cover more than 90% of the rose genome. An integrated map was constructed by incorporating the homologous parental linkage groups, resulting in seven linkage groups with a total length of 545 cM. The present linkage map is currently the most advanced map in rose with regard to marker density, genome coverage and with robust markers, giving good perspectives for QTL mapping and marker-assisted breeding in rose. The SSR markers, together with RFLP markers, provide good anchor points for future map alignment studies in rose and related species. Codominantly scored AFLP markers were helpful in the integration of the parental maps.

Chromosome Mapping↗

ROSE elements occur in disparate rhizobia and are functionally interchangeable between species.

Expression of at least ten genes in Bradyrhizobium japonicum, seven of which code for small heat shock proteins (sHsps), is under the control of ROSE (repression of heat shock gene expression). This negatively cis-acting DNA element confers temperature control to a sigma(70)-type promoter. Here, we show that ROSE elements are not restricted to B. japonicum but are also present in Bradyrhizobium sp. (Parasponia), Rhizobium sp. strain NGR234 and Mesorhizobium loti. An overall alignment of all ROSE sequences reveals a highly conserved and probably functionally important region towards the 3'-end of the element. Moreover, we provide genetic evidence for the previously proposed presence of multiple sHsps in these organisms. Primer-extension data of five newly identified ROSE-associated operons show that transcription is repressed at low temperatures and induced after a temperature upshift. Translational ROSE-hsp'-'lacZ fusions of Bradyrhizobium sp. (Parasponia) and Rhizobium sp. strain NGR234 integrated into the chromosome of B. japonicum were heat-responsive. The functionality of these heterologous ROSE elements hints at a common regulatory principle conserved in various rhizobia.

Bacterial Proteins↗

The association of Rose questionnaire angina pectoris and coronary calcification in a general population: the Rotterdam Coronary Calcification Study.

PURPOSE: The Rose questionnaire is a standardized method of measuring angina pectoris in general populations. Electron-beam computed tomography (CT) is a non-invasive technique to quantify the amount of coronary calcification. In a population-based study, we investigated the association between Rose questionnaire angina pectoris and coronary calcification. METHODS: The Rotterdam Coronary Calcification Study is embedded in the Rotterdam Study, a population-based study in subjects aged > or = 55 years. Participants of the Rotterdam Coronary Calcification Study underwent an electron-beam CT scan between 1997 and 2000. Coronary calcification was quantified in a calcium score according to Agatston's method. Rose questionnaire angina pectoris was assessed during a home interview. The first 2013 participants were included in the present analyses. RESULTS: In men, the presence of Rose questionnaire angina pectoris was associated with a 12.9-fold (95% confidence interval: 3.8-43.7) increased risk of a calcium score >1000 (reference: calcium score 0-100). The corresponding relative risk in women was 4.8 (2.0-11.3). Similar results were found when we computed sex-specific quartiles of the calcium score. CONCLUSIONS: Rose questionnaire angina pectoris is strongly associated with the amount of coronary calcification. Rose questionnaire angina pectoris corresponds better with the amount of coronary calcification in men than in women.

Aged↗

Development of a headspace-solid phase micro extraction method to monitor changes in volatile profile of rose (Rosa hybrida, cv David Austin) petals during processing.

In the present study, headspace solid phase microextraction combined to capillary gas chromatography (HS-SPME-GC) has been applied for the determination of changes in the volatile profile of rose petals (Rosa hybrida, cvs David Austin) following processing (heat treatment and addition as an ingredient to a food product--for example yoghurt). Four SPME fibres at two sampling temperatures (40 and 60 degrees C) with a sampling time of 30 min were examined. Volatile profiles were detected either by FID or/and by olfactometry (ODP-II, Gerstel). Fibre testing was performed using raw rose petals for sampling temperature selection and an 18 characteristic rose volatile standard mixture in water was used to compare fibre performances at the sampling temperature of 60 degrees C. Polydimethylsiloxane-divinylbenzene (PDMS-DVB) fibre at the sampling temperature of 60 degrees C was the most suitable to sample the rose alcohols phenyl ethanol, citronellol, nerol, geraniol and eugenol, as assessed by GC-olfactometry, not only from raw petals, but also from processed rose petals and the food product. PDMS-DVB fibre also showed a desired low affinity to volatiles from yoghurt, which reduces the influence of food matrix on the volatile profile. The method was linear over two orders of magnitude and had satisfactory repeatability, with limits of detection for the rose alcohols ranging from <1 to 10 ng/ml concentration levels.

Chromatography, Gas↗

Anticonflict effects of rose oil and identification of its active constituents.

The present study investigates the pharmacologically active constituents of rose oil, which possesses anti-conflict effects. Analysis using GC/MS revealed that rose oil contains 9 substances that were identified as myrcene, benzyl alcohol, 2-phenethyl alcohol, citronellol, geraniol, citronellyl acetate, eugenol, geranyl acetate and methyl eugenol. We examined the effects of each of these substances using the Geller and Vogel conflict tests in ICR mice. Myrcene, benzyl alcohol and citronellyl acetate did not produce any effects in either tests. Geranyl acetate and methyl eugenol produced no effect in the Geller conflict test. Geraniol and eugenol decreased the response rate during the safe period of the Geller conflict test, but did not affect the response rate during the alarm period. In contrast, 2-phenethyl alcohol and citronellol, like rose oil, produced an increasing effect on the response rate during the alarm period in the Geller conflict test. In addition, both chemicals increased the number of electric shocks mice received in the Vogel conflict test in a manner similar to that of rose oil. Given that 2-phenethyl alcohol and citronellol produced the same anti-conflict effects in both tests as rose oil, we concluded that they are the pharmacologically active constituents of anti-anxiety-like effect of rose oil.

Animals↗

A case of anaphylaxis due to rose pollen ingestion.

BACKGROUND: Recent publications have demonstrated that in Güneykent (Turkey), a rose-cultivating area, some workers experience various allergic reactions due to contact with rose or its pollen. CASE REPORT: A 47-year-old man, originally from Güneykent although he no longer lived there, suffered perennial rhinitis, asthma and rhinoconjunctivitis in spring related to sensitization to dust mites and various pollens, respectively. On several occasions, he had presented oral and cutaneous symptoms and angioedema due to contact with rose or intake of honey or other products. Finally, he had an anaphylactic reaction on drinking dew collected in the tulips of a rose that possibly contained rose pollen. METHODS: Specific IgE was evaluated with Rosa rugosa extract and was 30.3 IU (class IV). Sensitization to other pollens and to mites was also confirmed. Oral provocation test was not deemed ethical. CONCLUSION: This may be the first reported case of anaphylactic reaction due to rose pollen ingestion.

Anaphylaxis↗

O-methyltransferases involved in the biosynthesis of volatile phenolic derivatives in rose petals.

Rose (Rosa hybrida) flowers produce and emit a diverse array of volatiles, characteristic to their unique scent. One of the most prominent compounds in the floral volatiles of many rose varieties is the methoxylated phenolic derivative 3,5-dimethoxytoluene (orcinol dimethyl ether). Cell-free extracts derived from developing rose petals displayed O-methyltransferase (OMT) activities toward several phenolic substrates, including 3,5-dihydroxytoluene (orcinol), 3-methoxy,5-hydroxytoluene (orcinol monomethyl ether), 1-methoxy, 2-hydroxy benezene (guaiacol), and eugenol. The activity was most prominent in rose cv Golden Gate, a variety that produces relatively high levels of orcinol dimethyl ether, as compared with rose cv Fragrant Cloud, an otherwise scented variety but which emits almost no orcinol dimethyl ether. Using a functional genomics approach, we have identified and characterized two closely related cDNAs from a rose petal library that each encode a protein capable of methylating the penultimate and immediate precursors (orcinol and orcinol monomethyl ether, respectively) to give the final orcinol dimethyl ether product. The enzymes, designated orcinol OMTs (OOMT1 and OOMT2), are closely related to other plant methyltransferases whose substrates range from isoflavones to phenylpropenes. The peak in the levels of OOMT1 and OOMT2 transcripts in the flowers coincides with peak OMT activity and with the emission of orcinol dimethyl ether.

Amino Acid Sequence↗

A population study of the long-term consequences of Rose angina: 20-year follow-up of the Renfrew-Paisley study.

OBJECTIVE: To examine the long-term cardiovascular consequences of angina in a large epidemiological study. DESIGN: Prospective cohort study conducted between 1972 and 1976 with 20 years of follow-up (the Renfrew-Paisley Study). SETTING: Renfrew and Paisley, West Scotland, UK. PARTICIPANTS: 7048 men and 8354 women aged 45-64 years who underwent comprehensive cardiovascular screening at baseline, including the Rose Angina Questionnaire and electrocardiography (ECG). MAIN OUTCOME MEASURES: All deaths and hospitalisations for cardiovascular reasons occurring over the subsequent 20 years, according to the baseline Rose angina score and baseline ECG. RESULTS: At baseline, 669 (9.5%) men and 799 (9.6%) women had angina on Rose Angina Questionnaire. All-cause mortality for those with Rose angina was 67.7% in men and 43.3% in women at 20 years compared with 45.4% and 30.4%, respectively, in those without angina (p<0.001). Values are expressed as hazards ratio (HR) (95% confidence interval (CI). In a multivariate analysis, men with Rose angina had an increased risk of cardiovascular death or hospitalisation (1.49 (1.33 to 1.66), myocardial infarction (1.63 (1.41 to 1.85)) or heart failure (1.54 (1.13 to 2.10)) compared with men without angina. The corresponding HR (95% CI) for women were 1.38 (1.23 to 1.55), 1.56 (1.31 to 1.85) and 1.92 (1.44 to 2.56). An abnormality on the electrocardiogram (ECG) increased risk further, and both angina and an abnormality on the ECG increased risk most of all compared with those with neither angina nor ischaemic changes on the ECG. Compared with men, women with Rose angina were less likely to have a cardiovascular event (0.54 (0.46 to 0.64)) or myocardial infarction (0.44 (0.35 to 0.56)), although there was no sex difference in the risk of stroke (1.11 (0.75 to 1.65)), atrial fibrillation (0.84 (0.38 to 1.87)) or heart failure (0.79 (0.51 to 1.21)). CONCLUSIONS: Angina in middle age substantially increases the risk of death, myocardial infarction, heart failure and other cardiovascular events.

Age Distribution↗

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↗

Pharmacological studies of arrhythmias induced by rose bengal photoactivation.

Singlet oxygen and superoxide production by rose bengal photoactivation leads to rapid electrophysiological changes and arrhythmias. To investigate which intermediate is causative and to probe possible mechanisms, hearts (n = at least 6/group) were perfused aerobically for 10 min without rose bengal followed by 5 min with rose bengal before illumination for 20 min. In controls, all or most hearts exhibited ventricular premature beats, ventricular tachycardia, and complete atrioventricular block. Most antioxidants tested had no protective effect; histidine, however, significantly delayed the onset of electrocardiographic (ECG) changes. In further studies, two antiarrhythmic agents (quinidine and verapamil) had no little protective effect, whereas R56865 significantly delayed the onset of ECG changes and reduced the incidence of arrhythmias. However, spectrophotometric and laser pulse radiolysis studies showed that this apparent protective effect might have resulted from an interaction between R56865 and the rose bengal molecule, leading to a reduction in singlet oxygen production. In conclusion, the electrophysiological changes induced by rose bengal photoactivation are likely to be due to singlet oxygen; antiarrhythmic drugs appear to be unable to protect against the injury unless there is some interaction with the photoactivation process.

Animals↗

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↗