More names for depression: is a rose a rose?
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Molecular investigation of the progenitors, origin and domestication patterns of diploid Chinese old garden roses.
BACKGROUND AND AIMS: Chinese old garden roses are major contributors to the genetic development of modern roses. The RoKSN gene is associated with continuous flowering in roses and is proposed to have originated from Chinese wild roses. However, the wild roses that are implicated in the breeding of Chinese old garden roses and the origin of the RoKSN locus remain unidentified. We collected 25 of the most renowned and classic diploid Chinese old garden roses along with all related wild roses from East Asia. These roses were analysed with the aim of identifying the wild species that contributed to the genetic composition of Chinese old garden roses. In addition, we aimed to infer the geographical origin of the RoKSN gene and to develop a schematic overview of hybrid domestication of Chinese old garden roses. METHODS: We compared the haplotypes of internal transcribed spacers (nrITS), six nuclear single-copy genes and three chloroplast genes between Chinese old garden roses and wild roses. Additionally, we assessed genetic organization using 21 expressed sequence tag-simple sequence repeats to identify potential donor species that contributed to the emergence of these cultivars. Primers were designed for RoKSN to allow comparison of the gene across the entire distribution range of Rosa sect. Chinenses. KEY RESULTS: Our findings confirmed that the majority of rose cultivars are descendants of early hybridization events. Rosa chinensis var. spontanea, R. odorata var. gigantea and R. multiflora var. cathayensis were the primary donors for the 25 cultivar roses. Chinese old garden roses were categorized into four groups. Ten cultivars were hybrids between R. chinensis var. spontanea and R. multiflora var. cathayensis, thereby forming the 'Old Blush' group. Five cultivars were hybrids between 'Old Blush' and the R. kwangtungensis species complex, thereby forming the 'Slater's crimson' group. Six cultivars were hybrids between 'Old Blush' and R. odorata var. gigantea, thereby forming the 'Tea Rose' group, and three cultivars were hybrids that evolved from more than three donors. Moreover, we observed relatively close genetic proximity among Chinese old garden roses with an identical RoKSN-copia gene that is responsible for continuous flowering, which indicates a single origin for this retrotransposon-containing allele. Additionally, we determined that the haplotypes of the RoKSN-copia gene predominantly occurred in the Sichuan Basin region. In contrast, R. chinensis cultivated in the Ya'an region showed no markers of hybridization and displayed a genetic composition that was close to that of the wild species R. chinensis var. spontanea. This cultivar may represent the earliest mutated individual that bears the RoKSN-copia gene and may have served as a bridge from wild species to continuous-flowering old rose cultivars. CONCLUSIONS: The study provides crucial evidence that elucidates the origin of cultivated roses and lays the groundwork for further analysis of the breeding history of Chinese old garden roses using genomic data.
Domestication↗
Allergy symptoms and IgE immune response to rose: an occupational and an environmental disease.
BACKGROUND: Turkey is one of the four major producers of rose. Cultivation of rose is the main economic activity of many villagers in the Lakes region. Rose allergy has not been reported before. We investigated the prevalence of allergy symptoms and specific-IgE immune response due to rose in villagers who had been cultivating rose in Güneykent village in the Lakes region. METHODS: A screening questionnaire including respiratory and allergic symptoms was administered to 290 villagers. The investigation team visited and studied 75 randomly selected villagers with an interviewer-administered questionnaire, pulmonary function testing, skin prick testing and serum total IgE. Specific IgE against Rosa rugosa was measured in 41 villagers. RESULTS: Villagers reported asthma/allergy symptoms outside the rose season (17.6%), during the rose season (6.2%), and both during the rose season and outside the rose season (whole the year) (17.6%). Atopy and specific IgE against Rosa rugosa were detected in 12 (19%), and 8 (19.5%) of the villagers tested. Villagers who had symptoms whole the year reported wheeze more frequently than those who reported symptoms only outside the rose season (41.2% vs 11.1%). CONCLUSIONS: IgE-mediated allergy could occur due to rose and/or its pollen. Symptoms mainly affect the upper airways. Further studies in the rose handlers, florists and workers of the rose industry would help elucidate the occurrence of rose allergy.
Adult↗
Increased mortality among women with Rose angina who have not presented with ischaemic heart disease.
BACKGROUND: Little is known about the clinical importance of disease that is not presented to healthcare services. AIM: To determine the 5-year mortality among those with angina symptoms, known or not known by their general practitioner (GP) to have ischaemic heart disease (IHD). DESIGN: A prospective cohort study. SETTING: The study was conducted in the United Kingdom as part of the Royal College of General Practitioners' Oral Contraception Study. METHOD: In 1994-1995 women (n = 11,797) still under GP observation were sent a questionnaire that inquired about their smoking habits, other lifestyle issues, general health, and selected symptoms (including chest pain, assessed using the Rose angina questionnaire). The main outcome measure was the chances (odds) of dying during the next 5 years, among those with and without exertional chest pain, Rose angina or Rose myocardial infarction (MI), stratified by documented history of IHD. RESULTS: Overall, the lifetime prevalence of any exertional chest pain was 10.1% (95% confidence interval [CI] = 9.5 to 10.8); grade I Rose angina was 6.1% (95% CI = 5.6 to 6.6); grade II Rose angina was 1.3% (95% CI = 1.1 to 1.6); and Rose MI was 4.4% (95% CI = 4.0 to 4.9). The prevalence of each condition tended to increase with age, social class, parity, body mass index, and documented history of IHD. The proportion of women documented as having IHD was 23% among those with any exertional chest pain, 21.7% for grade I Rose angina, 37.7% for grade II Rose angina, and 31.4% for Rose MI. Compared to women without Rose angina, significantly higher odds ratios for all-cause mortality were observed among women with grade I Rose angina and no documented history of IHD (adjusted odds ratio [AOR] = 1.71, 95% CI = 1.05 to 2.79); those with grade II Rose angina and documented IHD (AOR = 3.94, 95% CI = 1.58 to 9.83); and women with grade II Rose angina and no documented history of IHD (AOR = 3.35, 95% CI = 1.47 to 7.62). CONCLUSIONS: Women with angina symptoms that have not been documented by their GP appear to have an increased risk of future mortality. Research is needed to determine the best way of identifying and managing these individuals.
Adult↗
Mucin characteristics of human corneal-limbal epithelial cells that exclude the rose bengal anionic dye.
PURPOSE: Rose bengal is an organic anionic dye used to assess damage of the ocular surface epithelium in ocular surface disease. It has been proposed that mucins have a protective role, preventing rose bengal staining of normal ocular surface epithelial cells. The current study was undertaken to evaluate rose bengal staining in a human corneal-limbal epithelial (HCLE) cell line known to produce and glycosylate membrane-associated mucins. METHODS: HCLE cells were grown to confluence in serum-free medium and switched to DMEM/F12 with 10% serum to promote differentiation. Immunolocalization of the membrane-associated mucins MUC1 and MUC16 and the T-antigen carbohydrate epitope was performed with the monoclonal antibodies HMFG-2 and OC125 and jacalin lectin, respectively. To assess dye uptake, cultures were incubated for 5 minutes with 0.1% rose bengal and photographed. To determine whether exclusion of negatively charged rose bengal requires a negative charge at the cell surface, cells were incubated with fluoresceinated cationized ferritin. The effect of hyperosmotic stress on rose bengal staining in vitro was evaluated by increasing the ion concentration (Ca+2 and Mg+2) in the rose bengal uptake assay. RESULTS: The cytoplasm and nucleus of confluent HCLE cells cultured in media without serum, lacking the expression of MUC16 but not MUC1, as well as human corneal fibroblasts, which do not express mucins, stained with rose bengal. Culture of HCLE cells in medium containing serum resulted in the formation of islands of stratified cells that excluded rose bengal. Apical cells of the stratified islands produced MUC16 and the T-antigen carbohydrate epitope on their apical surfaces. Colocalization experiments demonstrated that fluoresceinated cationized ferritin did not bind to these stratified cells, indicating that rose bengal is excluded from cells that lack negative charges. Increasing the amounts of divalent cations in the media reduced the cellular area protected against rose bengal uptake. CONCLUSIONS: These results indicate that stratification and differentiation of corneal epithelial cells, as measured by the capacity to produce the membrane-associated mucin MUC16 and the mucin-associated T-antigen carbohydrate on their apical surfaces provide protection against rose bengal penetrance in vitro and suggest a role for membrane-associated mucins and their oligosaccharides in the protection of ocular surface epithelia.
Antibodies, Monoclonal↗
Rose bengal inhibits herpes simplex virus replication in vero and human corneal epithelial cells in vitro.
Rose bengal dye is thought to highlight corneal lesions induced by herpes simplex virus type 1 (HSV-1) by virtue of its binding to dead or dying HSV-1-infected epithelial cells. However, whether rose bengal binds specifically to damaged versus normal corneal epithelial cells is unclear. To determine the binding properties of rose bengal, the authors compared binding of the dye to HSV-1-infected and uninfected cells, determined the cellular binding sites of the dye, and investigated the effects of rose bengal on HSV-1 replication in dye-treated cells in vitro. Spectrophotometric analysis revealed that uninfected and infected Vero cells bound equivalent amounts of dye at several times post inoculation, indicating that rose bengal does not preferentially bind to HSV-1-infected cells. By light microscopy, rose bengal was found to bind to the cell nuclei and perinuclear region of human corneal epithelial cells (HCEC) and Vero cells. Pretreatment of Vero and HCEC with different concentrations of rose bengal and exposure to 148 microW/cm2 of white light for 2 min reduced the ability of both cell types to support HSV-1 replication. Vero cells, in the absence of rose bengal, supported HSV-1 replication, whereas pre-treatment with 0.05% rose bengal reduced the yield of HSV-1 by 99.99% (P less than 0.000001) and 1% rose bengal completely prevented HSV replication. HCEC supported HSV-1 replication in the absence of rose bengal, but pre-treatment with 1% or 0.05% rose bengal completely prevented HSV-1 replication (P less than 0.000001).(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Promoter-specific synthetic photoendonuclease: rose bengal-labeled T7 RNA polymerase.
We have constructed a synthetic photoendonuclease composed of T7 RNA polymerase linked to rose bengal. The promoter-specific polymerase confers site-specific binding, and the photosensitizer rose bengal allows light-induced DNA cleavage. Using a gentle labeling procedure, we find that the polymerase can be labeled with 1-30 rose bengals. Polymerase labeled to about 8 rose bengals per molecule retains the same efficiency and specificity of binding to promoter-containing DNA as unlabeled polymerase. At this level of rose bengal substitution, the synthetic endonuclease, in the presence of visible light, specifically cleaves linear or supercoiled DNA containing a T7 promoter. It induces frank single-strand breaks, rather than labile sites convertible to breaks upon additional treatments. Neither the free rose bengal moiety not bonded to polymerase nor the free (not bound to DNA) rose bengal-substituted polymerase cleaves DNA. Although rose bengal is an efficient generator of singlet oxygen, depletion of oxygen from reaction mixtures increases the cleavage rate. This indicates that singlet oxygen cleavage is not a major mechanism of DNA nicking by the synthetic endonuclease. At higher levels of rose bengal substitution, the labeled polymerase shows decreased binding efficiency and increased nonspecific binding to DNA without a T7 promoter; the specificity of DNA cleavage also decreases. These results indicate that the site specificity of rose bengal photocleavage by the synthetic endonuclease results from specific binding of the polymerase, and thus rose bengal photonicking reflects polymerase binding.
Binding Sites↗
Skeletal sarcoplasmic reticulum dysfunction induced by reactive oxygen intermediates derived from photoactivated rose bengal.
We investigated the role of reactive oxygen intermediates generated from photoactivation of xanthene dye rose bengal on skeletal sarcoplasmic reticulum (SR) function, which plays a major role in the regulation of intracellular Ca++ and thereby in the generation of force. We used SR microsomes of canine masseter muscle as a model system in which to explore the effect of oxidation by determining oxalate-supported Ca++ uptake, Ca++, Mg++-adenosine triphosphatase (Ca++-ATPase) activity and Ca++ permeability of the SR vesicles. Skeletal SR vesicles exposed to rose bengal (50 nM) illuminated at 560 nm resulted in significant inhibition of Ca++ uptake velocity and Ca++-ATPase activity and in stimulation of Ca++ permeability. The observed effect afforded by illuminated rose bengal was dependent on intensity of light. Most reactive oxygen species scavengers tested had no protective effect; histidine (a powerful quenching agent for singlet oxygen), however, significantly protected the effect of illuminated rose bengal on Ca++ uptake velocity and Ca++-ATPase activity. The illumination of rose bengal also caused histidine-inhibitable loss of total sulfhydryl groups of SR. The increased Ca++ permeability elicited by illuminated rose bengal was blunted by a cocktail of histidine-catalase, but not by histidine alone. Generation of reactive oxygen species (singlet oxygen, superoxide and hydroxyl radical) from photoactivation of rose bengal was studied by electron spin resonance spectroscopy by use of the spin trap 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) and 2,2,6,6-tetramethylpiperidine (TEMP). We found that illumination of rose bengal formed a 1:2:2:1 quartet, characteristic of the hydroxyl radical-DMPO spin adduct, which was effectively blunted by hydroxyl radical scavenger, dimethyl sulfoxide, and by superoxide scavenger, superoxide dismutase. The results of electron spin resonance study also showed that singlet oxygen was produced by photoactivation of rose bengal was detected as singlet oxygen-TEMP product (TEMPO); 2,2,6,6-tetramethylpiperidine-N-oxyl). The formation of TEMPO signal was strongly inhibited by histidine. Similarly, we could detect hydrogen peroxide production from illuminated rose bengal. It is suggested that photoactivation of rose bengal generated singlet oxygen, superoxide, hydrogen peroxide and hydroxyl radical, and the data obtained from the present study indicate that singlet oxygen, rather than superoxide, hydrogen peroxide and hydroxyl radical, to be the active agent in the Ca++ transport system of SR; the observed effect of singlet oxygen may be due to sulfhydryl group oxidation. Our results are also consistent with the view that singlet oxygen does not appear to be an exclusive species that increases Ca++ permeability of SR vesicles, but the increased Ca++ permeability may be caused in part by hydrogen peroxide as well as singlet oxygen.
Animals↗
Angina pectoris by Rose questionnaire does not predict cardiovascular disease in treated hypertensive patients.
OBJECTIVE: To determine the predictive value of angina pectoris diagnosed by Rose questionnaire for cardiovascular disease among treated hypertensives. METHODS: The cardiovascular experience of 4093 patients who had no history of cardiovascular disease and had been administered the Rose questionnaire for angina in a worksite treatment program was evaluated. RESULTS: Among 2659 men and 1434 women of similar age (53 versus 54 years), the race distribution was 44 versus 31% whites, 27 versus 41% blacks and 29 versus 28% Hispanics. Overall, the prevalence of angina by Rose questionnaire in women (15%) was twice that in men (7%) in all three races, with Hispanics having the highest (20 versus 10%) prevalence. Those with angina (Rose-plus) and those without (Rose-minus) had similar initial and final blood pressures. In 4.0 years of average follow-up study, the crude incidence rates (per 1000 person-years) of the recorded 120 myocardial infarctions and 35 strokes did not differ significantly between Rose-plus and Rose-minus patients, except for myocardial infarction in Hispanic men (20.5 versus 5.9). When myocardial infarction incidence was adjusted for age within each sex-race subgroup, only Rose-plus Hispanic men had a significantly greater relative risk with Rose-minus as referent (relative risk 3.13, 95% confidence interval 1.31-7.50). Overall, in the Cox proportional hazards regression model, angina by Rose questionnaire was not predictive of myocardial infarction after accounting for other recognized risk factors. CONCLUSIONS: The present data suggest that the Rose questionnaire as a diagnostic tool for angina is not predictive of subsequent clinical events among treated hypertensive patients.
Adult↗
The performance of the Rose angina questionnaire in South Asian and European origin populations: a comparative study in Newcastle, UK.
BACKGROUND: The Rose angina questionnaire has been extensively used in different cultural settings, but may not perform consistently in different ethnic groups. We set out to assess the performance of the Rose angina questionnaire in UK South Asians compared with Europeans. METHODS: Data on major ECG abnormalities, possible or definite Rose questionnaire angina and diagnosed angina were collected from an age- and sex-stratified random sample of 1509 adults from European, Indian, Pakistani and Bangladeshi ethnic groups. RESULTS: The ECG abnormalities were commoner in South Asians than Europeans (6% versus 2% in men). The prevalence in both South Asian and European men of possible Rose angina and diagnosed angina was 18% and 8%, respectively, but definite Rose angina was less common in South Asians (3% versus 6%). Definite Rose angina showed lower sensitivity for other measures in South Asians than in Europeans: sensitivity for a doctor's diagnosis was 21% in South Asian and 37% in European men. For possible Rose angina, the corresponding figures were 81% and 84%. Similar patterns were seen in women. CONCLUSIONS: The performance of the Rose angina questionnaire was sufficiently inconsistent to warrant further work to achieve greater cross-cultural validity. Possible Rose angina performed more consistently across ethnic groups than definite Rose angina and pending further validation studies may be the most appropriate form to use.
Angina Pectoris↗
Performance of the WHO Rose angina questionnaire in post-menopausal women: are all of the questions necessary?
OBJECTIVE: To assess the performance of a shortened version of the Rose angina questionnaire focusing on exertional chest pain. METHODS: Cross sectional analysis of 3987 women aged 60 to 79 years from 23 British towns. The performances of definite Rose angina (using data from the full Rose angina questionnaire) and exertional chest pain (using data from a subset of three questions from the Rose angina questionnaire) were assessed against a medical record of angina. RESULTS: The sensitivity (the proportion with a medical record of angina who were identified as having angina by the questionnaire) was 29.9% (95% confidence intervals 25.7% to 34.4%) comparing definite Rose angina to any medical record of angina since 1978 and 50.7% (45.9% to 55.5%) comparing exertional chest pain to any medical record diagnosis of angina. The positive predictive values of both questionnaires were similar. When the two questionnaires were compared with a gold standard of a primary care consultation for angina symptoms within the past five years the sensitivity of definite Rose angina was 33.0% (26.9% to 39.6%) and that of exertional chest pain was 51.8% (45.1% to 58.5%). Although the sensitivity of both versions of the questionnaire was greater in those aged 60-69 years compared with those aged 70-79 years, it remained higher in the exertional chest pain version of the questionnaire than for definite Rose angina based on the full version of the questionnaire in both age groups. Performance of either version of the questionnaire was not affected by occupational social class. CONCLUSIONS: With respect to identifying women with a medical diagnosis of angina or those presenting to primary care with anginal symptoms, these results suggest that a shortened version of the Rose angina questionnaire focusing on exertional chest pain performs better than the full version. Other studies suggest that exertional chest pain is the crucial element of the Rose angina questionnaire with respect to predicting future coronary events. It is concluded that using a shortened version of the Rose angina questionnaire is adequate in epidemiological studies.
Aged↗
Patient tolerance and ocular surface staining characteristics of lissamine green versus rose bengal.
PURPOSE: To determine patient tolerance and ocular surface staining characteristics of 1% lissamine green versus 1% rose bengal solutions in patients with dry eye and in those with normal ocular surfaces by comparison of subjective sensation and objective staining scores. METHODS: Twelve patients with keratoconjunctivitis sicca and eight subjects with no ocular surface disease recorded their symptoms on a graded scale (0-5) and the duration of symptoms after instillation of one drop of 1% lissamine green and 1% rose bengal on two different occasions in different order. Lissamine green and rose bengal staining patterns were recorded on a graded scale (0-4) by one of the authors. RESULTS: For patients with keratoconjunctivitis sicca mean sensation score with lissamine green (2.42) was significantly lower (P = 0.00006) than with rose bengal (4.58). The duration of symptoms was significantly longer (P = 0.0007) after rose bengal instillation as well. For subjects with no ocular surface disease, mean sensation score with lissamine green (1.375) was significantly lower (P = 0.01) than with rose bengal (2.5). In this group, duration of symptoms also was significantly longer (P = 0.001) after rose bengal instillation. In both groups, there was no difference in objective staining scores. CONCLUSION: Lissamine green is better tolerated than rose bengal by patients and is equally as effective as rose bengal in evaluating the ocular surface in keratoconjunctivitis sicca.
Adult↗
Interaction between rose bengal and different protein components.
Bindings of rose bengal to several proteins were determined by Sephadex G-75 chromatography. Their respective blocking effect against dye uptake was demonstrated in an assay using a rabbit corneal epithelial cell layer. The total binding capacity of nonmucin proteins was measured using fluorometry and Scatchard analysis. The results showed that albumin, lactoferrin, transferrin, and lysozyme could--but serum prealbumin, IgA, carboxymethyl cellulose (CMC), and Sepharose 4B-purified porcine stomach mucin (PSM) could not--bind rose bengal. Lysozyme formed precipitates with rose bengal. Sufficient concentrations of albumin, lactoferrin, transferrin, or lysozyme premixed with rose bengal could block dye uptake by cells, but IgA and serum prealbumin could not. Premixed PSM was not as effective as precoated PSM in blocking dye uptake. The dissociation constant (Kd) was 1.2 x 10(-7) M, 3.6 x 10(-7) M, 3.9 x 10(-7) M, and 1.6 x 10(-6) M for albumin, transferrin, lactoferrin, and lysozyme, respectively. Based on these values, the total maximal binding capacity of nonmucin proteins in normal 7-microliters tears was extrapolated to be 0.249 micrograms rose bengal, which is too small to explain the negative staining of rose bengal on the normal ocular surface. Rose bengal, but not fluorescein, could interact with carbohydrate-containing Sephadex, CMC, and PSM to slow down its elution via Sephadex column chromatography. Therefore, the normal negative staining to rose bengal might be caused by the blocking effect of preocular mucus tear layer, which serves as a diffusion barrier. Rose bengal remains a unique dye for detecting the protective function of the preocular mucus tear.
Animals↗
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↗
PCR assessment of HSV-1 corneal infection in animals treated with rose bengal and lissamine green B.
PURPOSE: In vivo, the ophthalmic dye rose bengal displays profound antiviral effects against herpes simplex virus (HSV)-1, thus limiting its utility in diagnosis of epithelial keratitis when used before viral culture is performed. In contrast, lissamine green B does not possess significant antiviral activity in vivo. To determine whether polymerase chain reaction (PCR) could successfully detect HSV-1 DNA in ocular samples that have been exposed to ophthalmic dyes, animal models were used to observe the presence of infectious HSV-1 and viral DNA in eyes treated with rose bengal or lissamine green B. METHODS: Animals were bilaterally infected with HSV-1 strain H129, and at daily intervals up to 16 days post infection (dpi) rose bengal or lissamine green B was instilled in the left eyes. The right eyes were not treated with dyes. Swabs of the dye-treated and untreated eyes were assayed by PCR for viral infectivity by culture and the presence of DNA specific for a fragment of the HSV-1 DNA polymerase gene. RESULTS: A statistically equivalent number of samples from lissamine green B-treated and untreated eyes were positive by both viral culture and PCR. In contrast, rose bengal significantly decreased the infectious virus present in ocular secretions. A total of 44% and 78% of the rose bengal-treated and untreated eye samples, respectively, were positive by culture from 1 through 16 dpi. PCR was more sensitive than culture for detection of HSV-1 in rose bengal-treated eyes, in that 74% of rose bengal-treated samples were positive by PCR compared with 44% that were positive by culture during the 16-day period studied. It was also noted that both rose bengal and lissamine green B treatments slightly prolonged the period during which viral DNA was detectable in ocular secretions by PCR, possibly because the singlet oxygen produced by these photoreactive dyes compromised ocular cellular, humoral, and nonspecific immune factors allowing viral DNA to persist for slightly longer periods. CONCLUSIONS: PCR can successfully detect HSV-1 DNA in ocular samples that are culture negative and contain rose bengal or lissamine green B. Visualization of ocular epithelial defects with lissamine green B does not interfere with detection of infectious virus or HSV-1 DNA.
Animals↗
Visual performance and comfort with the Rose K lens for keratoconus.
PURPOSE: The "Rose K Lens for Keratoconus" is a proprietary rigid contact lens design that has been reported and marketed as a lens that provides better visual acuity and comfort than other rigid contact lens designs for keratoconus. METHODS: Twenty patients with keratoconus who were wearing rigid contact lenses were refitted into the Rose K lenses. Visual acuity measurements were taken with habitual lenses at the baseline visit and with the Rose K lenses at the completion of the study. Questionnaires were used to assess vision-specific quality of life, contact lens comfort, and self-reported assessment of vision. RESULTS: There were no statistically significant changes in high- or low-contrast visual acuity with the Rose K lenses. There was statistically significant improvement in self-reported assessment of vision and self-reported assessment of comfort in the eyes with more advanced keratoconus. At the conclusion of the study, 72% of patients preferred the Rose K lenses over their habitual lenses, and 87% reported that they would continue wearing the Rose K lenses. CONCLUSIONS: There was no difference in the visual acuity with the Rose K lenses compared with the patients' habitual lenses. Subjective assessment of vision and comfort indicate a statistical improvement for more advanced keratoconus with the Rose K lens. We could not rule out a placebo effect as a source of subjective improvement in vision and comfort. Nevertheless, the successful fit rate and patient preference demonstrate the usefulness of the Rose K lens in clinical practice.
Adult↗
[The quantity and structure of the root-associated microbial complexes of two greenhouse rose cultivars].
The study of the root-associated microbial complexes of affected and healthy rose plants of two cultivars (Grand gala and Royal velvet) grown in a greenhouse showed that the biomass of eukaryotic microorganisms in the rhizoplane and rhizosphere of healthy rose plants and in the surrounding soil was considerably lower than in the same loci of affected plants. In contrast, the biomass of root-associated prokaryotic microorganisms was higher in the case of healthy than in the case of affected rose plants. The root-associated bacterial complexes of both affected and healthy rose plants were dominated by the genera Arthrobacter, Rhodococcus, and Myxobacterium and did not contain phytopathogenic bacteria. The root-associated fungal complex of healthy roses was dominated by fungi of the genus Trichoderma, whereas that of the affected rose plants was dominated by the species Aureobasidium microstictum. The affected cane cuttings and cankers occurring on affected canes were found to contain Coniothyrium fuckelii (the causal fungus of rose stem canker) and sclerotia of Botrytis cinerea (the causal fungus of gray rot). The micromycete complex of healthy rose plants was not so diverse as was the micromycete complex of affected rose plants.
Arthrobacter↗
What is actually stained by rose bengal?
It has been believed that 1% rose bengal does not stain normal, healthy cells but rather stains degenerated or dead cells and mucous strands. In contrast to this conventional knowledge, we discovered that both commercial additive-containing and additive-free rose bengal solutions stained four different types of healthy cultured cells, including rabbit corneal epithelial cells. Rose bengal staining was rapid, dose dependent, predominantly nuclear, and detectable with the naked eye at concentrations as low as 0.05% and 0.025% for the commercial additive-containing or additive-free solutions, respectively, and with the fluorescence microscope at a concentration of 0.001%. It is surprising to discover that rose bengal is not a vital dye; after staining, cells actually lost vitality, as evidenced by instant morphologic changes, subsequent loss of cellular motility, cell detachment, and cell death. Such an intrinsic toxic effect was augmented by light exposure. The rose bengal staining of live as well as detergent-treated (Triton X-100) cells could be blocked by such tear components as mucin and albumin, suggesting that normally negative rose bengal staining is due to the protective function of the preocular tear film, ie, staining is not dictated by lack of cell vitality. These data indicate that rose bengal staining ensues whenever there is poor protection of surface epithelium by the preocular tear film; this represents a new interpretation for rose bengal stains seen in various ocular surface disorders.
Animals↗