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PubMed2001

Treatment of superior limbic keratoconjunctivitis by application of autologous serum.

PURPOSE: To determine the efficacy of autologous serum drops in the treatment of superior limbic keratoconjunctivitis (SLK). METHODS: Twenty-two eyes of 11 patients were diagnosed with SLK. All eyes were treated with 20% diluted autologous serum eyedrops 10 times a day in addition to ongoing treatment of dry eye. Fluorescein and rose bengal staining scores, as well as subjective symptom gradings, were performed before and after 4 weeks of therapy. RESULTS: Nine of the 11 patients responded well to treatment (82%). The average rose bengal and fluorescein score improved ( p < 0.05), and there was subjective improvement. CONCLUSION: Autologous serum application can be used as an alternative mode of therapy in SLK.

Administration, Topical↗

PubMed2005

[Prevalence of dry eye in Hansen's disease patients from a colony hospital in Goiânia, Brazil].

PURPOSE: To determine the prevalence of dry eye in Hansen's disease patients from the "Hospital de Dermatologia Sanitária de Goiânia". METHODS: The sample of the present study included 70 patients with Hansen's disease, from the "Hospital de Dermatologia Sanitária de Goiânia", and 30 controls, from the "Fundação Banco de Olhos de Goiás", both located in Goiânia-GO, Brazil. Ophthalmologic examination and Schirmer I, break-up time (BUT) and rose Bengal staining tests were carried out for all patients in a single evaluation. For the diagnosis of dry eye the Schirmer I equal or less than 5 mm and rose Bengal grade II or III, in at least one eye were considered. RESULTS: Forty-four (63.0%) patients with Hansen's disease were male and 22 (73.3%) controls were female (p=0.001). The mean age of the patients with Hansen's disease was 61.1+/-12.5 years and in the control group, it was 55.7+/-9.6 years. Fifteen (21.4%) patients with Hansen's disease and four (13.3%) controls presented with dry eye diagnosis (p=0.429). The lepromatous type (74.2%) of Hansen's disease was the most prevalent and dry eye (66.7%) was more frequent in this clinical form of the disease. CONCLUSION: The dry eye disease was more prevalent in patients with Hansen's disease, although the difference was not statistically significant.

Adult↗

PubMed1996

[The development of shrimp blood agar for testing the hemolysis of shrimp's haemocyte by bacteria].

A new plating medium, shrimp blood agar, was developed by using the haemolymph of shrimp plus 200 ppm rose bengal as the substrate. The colorless shrimp haemocytes were dyed by rose bengal to red. On the present agar, the hemolytic bacteria strain may show a clear zone surrounding the bacterial colony. The result on hemolysis of 45 bacteria representing 12 genera isolated from aquaculture environments against shrimp blood agar and sheep blood agar was also evaluated. There are 11 strains with different results. The study showed that, with the aim of screening the hemolytic bacteria to shrimp, shrimp blood agar might reveal a relatively quick and accurate results than that of sheep blood agar.

Agar↗

PubMed1979

Absorbed dose estimation of some 99mTc-hepatobiliary agents.

The absorbed dose estimates of several organs after administration in humans of six new hepatobiliary agents (five 99mTc-Pyridoxal amino acid complexes and the 99mTc-HIDA) are presented in comparison with 131I-Rose-Bengal. The results indicated that the radiation doses absorbed by the total body and gonads as well as by the critical organ (upper part of the large intestine) are significantly lower than those of Rose Bengal. Therefore, the new 99mTc-agents can be safely applied to humans.

Adult↗

PubMed1994

The role of reactive oxygen species in the degradation of lindane and DDT.

We have studied the photodegradation reactions of lindane and DDT under sunlight (72 hr) or a solar simulator (2 hr) either directly or in presence of selected photodynamic sensitizers. Direct photolysis did not cause noticeable degradation of the pesticides (below 1% in both the cases). However, addition of benzophenone or rose bengal to the irradiation solution of lindane resulted in 88% and 47% degradation, respectively, under sunlight. Similar experiments carried out with DDT resulted in 89%, 50% and 32% degradation with benzophenone, rose bengal and methylene blue, respectively. All the photosensitizers used for the study have been shown to induce photooxidation reactions by generation of reactive oxygen species. These studies suggest that by employing certain naturally occurring photosensitizers with reactive oxygen generation potential under sunlight, an effective method for safe removal of pesticides and other environmentally persistent agents from the surface of earth can be developed.

Anthracenes↗

PubMed2002

Photodynamic action of some sensitizers by photooxidation of luminol.

We report the development of a novel simple experimental method which allows the comparison of new photosensitizers based on their production of reactive oxygen species. A high-performance liquid chromatography (HPLC) assay permits the monitoring of several substances (sensitizer, reactant and oxidized end product) simultaneously on a single chromatogram. Photoreactions were monitored throughout their course by the HPLC assay surveying the sensitizers' efficiency of singlet oxygen production by the oxidative decomposition of luminol. Several photosensitizers were tested: Rose Bengal, Methylene Blue, Protoporphyrin IX, Photosan III, Photofrin, Hypericin and Pseudohypericin. Additionally, photoreactions were monitored by a standard pO(2) detection system. The measurements of the two detection methods were strongly correlated. Rose Bengal proved to be the most efficient photosensitizer, clearly decreasing the luminol concentration and causing a corresponding increase in aminophthalic acid. Our experiments show that when factors necessary for photochemical reactions are absent or are blocked (antioxidants), no reaction can be detected.

Dihematoporphyrin Ether↗

PubMed1995

Experimental thrombosis model induced by free radicals. Application to aspirin and other different substances.

A large number of experimental studies suggests that oxygen free radicals play a major role in the pathogenesis of the myocardial lesions observed during the sequence ischemia-reperfusion. The purpose of this study was to determine whether oxygen free radicals can induce thrombosis. In so doing we have developed a new experimental thrombosis model. Reproducible focal thrombosis has been achieved by irradiating mesenteric arterioles of rat for variable time with green filtered light issuing from a mercury lamp after systemic injection of different rose bengal doses. The number of emboli that remove in the blood (N), the duration of total occlusion (T) and the number of emboli per minute were then measured. As control, no rose bengal administration was done and the vessels were exposed to the filtered light. In comparison with this control, results clearly showed that free radicals always induced thrombosis and the induced thrombus was mainly composed of platelets. In this new thrombosis model induced by free radicals antithrombotic drugs (aspirin, 200 mg/Kg, heparin, 2 mg/Kg) and antioxidants (vitamin C, 10 and 20 mg/Kg, allopurinol, 200 and 300 mg/Kg, vitamin E, 500 and 1000 mg/Kg) have been tested. Results have shown that only heparin and vitamin E had an antithrombotic effect on thrombus formation induced by free radicals. This model should be useful in studying the effects of different drugs and could lead to new treatment modalities for ischemic accident and other cardiovascular diseases.

Allopurinol↗

PubMed1995

The specificity and product of quenching singlet oxygen by 2,2,6,6-tetramethylpiperidine.

The specificity of 2,2,6,6-tetramethylpiperidine to singlet oxygen was shown using Rose Bengal as a singlet oxygen generator, and Xanthine-Xanthine Oxidase and KO2 as the sources for the superoxide radical. The highest concentration of produced-singlet oxygen occurred at 25% of O2 by Rose Bengal photosensitization. The linewidth of the EPR signal for photosensitized nitroxyl radical, increasing solvent polarity. Deuterated solvents enlarge the EPR signal intensity in a dose-dependent manner. No EPR signal increase was observed in xanthine-xanthine oxidase reaction or KO2 systems, indicating that TEMP does not react with the superoxide anion. Thus, reaction of TEMP with 1O2 is highly specific.

Cyclic N-Oxides↗

PubMed1992

The diagnostic power of the tests for tear gland related keratoconjunctivitis sicca.

Laboratory tear function tests, such as the lactoferrin and the lysozyme test, reflect the level of tear gland deficiency and are, therefore, very useful for the clinician in the diagnosis of keratoconjunctivitis sicca (KCS). Although these tests do have an excellent discriminatory ability, they should be used in addition to clinical tests, such as the Schirmer-1 test, the tear film break-up time (BUT) and the Rose bengal (Rb) staining test. Schirmer's test, the most commonly used clinical test, measures tear fluid production. The tear film BUT, on the other hand, is the only test which establishes tear film instability. The Rose bengal score is strongly related to secondary damage of the superficial epithelium of the cornea and conjunctiva in patients with KCS. Thus, combining the results of these various tests facilitates the diagnosis of KCS and also the differentiation between Sjögren's KCS and non-Sjögren's KCS.

Diagnosis, Differential↗

PubMed1988

Sodium hyaluronate and polyvinyl alcohol artificial tear preparations. A comparison in patients with keratoconjunctivitis sicca.

An unpreserved artificial tear substitute containing 0.1% sodium hyaluronate was compared with a preparation containing 1.4% polyvinyl alcohol and 0.5% chlorobutanol in a controlled, double-masked, randomized study in patients with moderately severe keratoconjunctivitis sicca. Patients were evaluated initially, at 1, 4, and 8 weeks. The dry-eye status was evaluated by means of tear-film osmolality, tear breakup time, rose bengal staining, Schirmer's test (without anesthesia), and ocular surface-impression cytology. In general, neither preparation was found to be superior to the other. In both study groups, the mean tear-film osmolality and rose bengal staining score improved over the eight-week study, but the degree of squamous metaplasia of the bulbar conjunctival surface, as shown by impression cytology, did not change significantly.

Clinical Trials as Topic↗

PubMed1978

99MTc-HIDA, a gallbladder imaging agent: experimental aspects.

The molecule N-(2,6-dimethyl-phenyl-carbamoyl-methyl)-iminodiacetic acid (HIDA), capable of chelating reduced 99mTc, was synthesized, characterized, labeled with 99mTc, and studied in experimental animals. The results indicated that the new 99mTc-radiopharmaceutical is rapidly cleared from the blood to the liver, then rapidly removed to the gallbladder and excreted into the duodenum through the common bile duct. A comparative kinetic study of 99mTc-HIDA and 131I-Rose Bengal performed in rabbits demonstrated that both radiopharmaceuticals had a similar blood clearance rate, but cleared at a different rate from liver to gallbladder. 99mTc-HIDA showed a faster accumulation in the gallbladder than 131I-Rose Bengal. These findings, combined with the advantage of the low acute toxicity of HIDA, were promising enough to encourage a further evaluation and clinical investigation of this new Tc-99m hepatobiliary agent.

Animals↗

PubMed2003

Ocular surface changes in keratoconjunctivitis sicca with silicone punctum plug occlusion.

PURPOSE: To evaluate histopathologic and clinical response to silicone plug insertion in dry eye patients. METHODS: Punctal plugs were placed in 32 eyes of 18 dry eye patients with aqueous deficiency who were on maximum medical therapy and who had Schirmer testing with topical anesthetic measuring less than 5 mm. Pre-treatment and post-treatment (6 weeks and 1 year) evaluations included temporal and inferior quadrant impression cytology examinations, Schirmer-1, BUT, corneal fluorescein and rose-bengal staining patterns and questionnaire scores. The cytology samples were graded according to the method described by Nelson. RESULTS: There was an improvement in subjective symptoms of patients 6 weeks after punctal occlusion. Corneal fluorescein and rose-bengal staining scores decreased from a mean of 5.33 +/- 1.74 to 1.37 +/- 1.48 and from 5.90 +/- 1.2 to 2.45 +/- 1.89 (p < 0.05) respectively. BUT values increased from 2.98 +/- 1.1 to 11.3 +/- 3.1 seconds (p < 0.05) at 6 weeks post-treatment. Increase in goblet cell density was observed after punctal occlusion at 6 weeks (p < 0.05) and at 1 year (p < 0.05) compared to the pre-treatment values. After occlusion, 3% of temporal specimens had Grade 0 squamous metaplasia, 63% had Grade 1, 34% had Grade 2; while 25% of inferior specimens had Grade 0, 56% had Grade 1, 19% had Grade 2 squamous metaplasia. CONCLUSIONS: Punctum plug occlusion in keratoconjunctivitis sicca provides improvement of tear film stability, ocular surface staining scores, conjunctival squamous metaplasia grades and goblet cell density. Increased ocular surface exposure to essential tear components with punctal occlusion may be important for the genesis of these changes.

Adult↗

PubMed2002

New photoactivators for multiphoton excited three-dimensional submicron cross-linking of proteins: bovine serum albumin and type 1 collagen.

We report the synthesis and optical characterization of two new photoactivators and demonstrate their use for multiphoton excited three-dimensional free-form fabrication with proteins. These reagents were developed with the goal of cross-linking Type 1 collagen. This cross-linking process produces structures on the micron and submicron size scales. A rose bengal diisopropyl amine derivative combines the classic photoactivator and co-initiator system into one molecule, reducing the reaction kinetics and increasing cross-linking efficiency. This derivative was successful at producing stable structures from collagen, whereas rose bengal alone was not effective. A benzophenone dimer connected by a flexible diamine tether was also synthesized. This activator has two photochemically reactive groups and is highly efficient in cross-linking bovine serum albumin and Type 1 collagen to form stable, robust structures. This approach is more flexible in terms of cross-linking a variety of proteins than by traditional benzophenone photochemistry. The photophysical properties vary greatly from that of benzophenone, with the appearance of a new, lower energy absorption band (lambda max approximately 370 nm in water) and broad, visible emission band (approximately 500 nm maximum). This absorption band is highly solvatochromic, suggesting it arises, at least in part, from a charge transfer interaction. Collagens are typically difficult to cross-link photochemically, and the results here suggest that these two new activators will be suitable for cross-linking other forms of collagen and additional proteins for biomedical applications such as the de novo assembly of biomimetic tissue scaffolds.

Benzophenones↗

PubMed1995

Adenovirus epithelial keratitis.

The authors sought to determine whether adenovirus could infect human corneal epithelium in vivo. They reviewed the medical records of six patients with adenovirus-positive viral corneal cultures who were examined at the Bascom Palmer Eye Institute between March 21, 1986, and December 31, 1992. The six patients with adenovirus-positive viral corneal cultures included one patient with dendritic epithelial keratitis, one with geographic epithelial ulceration, three with both geographic ulceration and contiguous dendrites, and one with heaped-up corneal epithelium but no ulceration. Four patients had rose bengal solution applied to their ocular surface, and in all four patients rose bengal uptake was seen at the epithelial edges of the dendrite or geographic ulceration in a manner characteristic of herpes simplex viral keratitis. Serotype determination of the isolates showed adenovirus type 3 (one patient), type 8 (three patients), type 19 (one patient), and indeterminate (one patient). Results of monoclonal antibody staining of cultures against herpes simplex virus (types 1 and 2) antigens was negative for all six cases. Adenovirus epithelial keratitis may result from infection of human corneal epithelium by the virus and rarely may mimic infection of corneal epithelium by herpes simplex virus.

Adenovirus Infections, Human↗

PubMed1976

Tryptophanyl and carboxylic acid residues in the active centre of glucoamylase I from Aspergillus niger.

The pH-dependence of the photo-oxidation of L-tryptophan, in the presence of Rose Bengal and Methylene Blue, has been investigated. True, initial rate constants were determined in order to circumvent errors due to secondary processes. Photo-oxidation of glycoamylase I from A. niger in the presence of Methylene Blue or Rose Bengal resulted in a pH-dependent loss of enzymic activity, which was analogous to the destruction of free L-tryptophan during photo-oxidation. The loss of enzymic activity was closely associated with the destruction of tryptophan residues in the enzyme. Significant protection of both enzymic activity and tryptophanyl residues in the enzyme molecule was achieved by performing the photo-oxidation in the presence of maltose, which is a substrate for the enzyme. The tryptophanyl residues of glucoamylase I, which had been inactivated by reaction of its carboxylic acid residues with glycine methyl ester in the presence of a water-soluble carbodi-imide, were also substantially protected by maltose. It is concluded that the active centre of glucoamylase I is a cleft lined with tryptophanyl residues that participate in the binding of the substrate. One or more carboxylic acid residues are involved in bond cleavage.

Aspergillus↗

PubMedDate not supplied

Sperm decondensation and semen parameters: utilization of a simple staining technique for the evaluation of human sperm decondensation.

The ability of spermatozoa to fertilize an oocyte depends on a sequence of events ending ultimately in the decondensation of the sperm chromatin on penetration of the oocyte. Knowledge of what percentage of sperm decondenses is useful, especially in patients where other functional tests and sperm quality fail to explain the reported poor in vitro fertilization (IVF) rates. The objective of this study was (1) to compare sperm decondensation induced by either SDS/EDTA or heparin with semen parameters (volume, concentration, motility and morphology), and (2) to evaluate the use of a simplified staining technique (Diff QuikR [DQ]) in comparison with the standard phase contrast method (Rose Bengal-[RB]). Randomly selected semen samples from 31 men attending an assisted reproductive programme were analysed for basic semen parameters and decondensation with SDS/EDTA and heparin. Two staining methods for the evaluation of decondensation were compared (phase contrast microscopy after Rose Bengal [RB] staining and light microscopy after Diff QuikR (DQ) staining). Moderate and grossly swollen sperm heads were recorded. Semen samples included both fertile and unfertile semen parameters. Sperm decondensation results showed poor to moderate correlations with semen parameters. The SDS/EDTA (DQ) (moderate forms) showed a significant negative correlation (r = -0.46) with seminal volume and a significant positive correlation (r = 0.41) with normal sperm morphology. The heparin (DQ) (moderate forms) decondensation showed a significant positive correlation with motility (r = 0.61) and sperm concentration (r = 0.43). The DQ method was preferred over the RB method due to its optical and storage advantage.(ABSTRACT TRUNCATED AT 250 WORDS)

Azure Stains↗

PubMed2006

The role of Yap1p and Skn7p-mediated oxidative stress response in the defence of Saccharomyces cerevisiae against singlet oxygen.

The production of the reactive oxygen species superoxide and hydrogen peroxide in Saccharomyces cerevisiae induces the expression of various defence genes involved in an oxidative stress response. Expression of many of these genes has been shown to be coordinated by two transcriptional regulators, Yap1p and Skn7p, either alone or in concert. Here, we investigated the role of the Yap1p and Skn7p-mediated stress response in the defence against singlet oxygen, a non-radical reactive oxygen species produced mainly by photosensitized reactions in illuminated cells. Both, a yap1 and skn7 mutant were highly sensitive to Rose Bengal, an exogenous photosensitizer producing singlet oxygen in the light. The expression of a Yap1p-dependent reporter gene was induced by increased singlet oxygen production, showing that singlet oxygen activates general oxidative stress response mechanisms required for the resistance against Rose Bengal treatment. This response was also slightly stimulated by light in the absence of the photosensitizer, possibly due to singlet oxygen production by endogenous photosensitizers. The expression pattern of four oxidative stress genes in a yap1, skn7 and wild-type strain and the sensitivity of the corresponding mutants exposed to different oxidative stress conditions proved a role of Yap1p and Skn7p in the defence against singlet oxygen. Similarities in the genetic responses against singlet oxygen and hydroperoxides suggest an overlap in the oxidative stress response against these reactive oxygen species.

DNA-Binding Proteins↗

PubMed2004

Dry eye syndrome in thyroid-associated ophthalmopathy: lacrimal expression of TSH receptor suggests involvement of TSHR-specific autoantibodies.

PURPOSE: We evaluated the relationship between ocular surface damage, elevated lid aperture/impaired Bell's phenomenon and reduced tear production in thyroid-associated ophthalmopathy (TAO). Suspecting a possible role of autoantibodies specific for TSH receptor (TSHR), we further investigated TSHR expression in the healthy lacrimal gland (LG). METHODS: A total of 48 patients with active TAO and 26 controls were examined for basal tear secretion, Rose Bengal and fluorescein staining, impression cytology (IPC), break-up time (BUT), and blinking (lid width, lid closure, ocular surface, upward excursion). Healthy LGs were investigated immunohistochemically for expression of TSHR. RESULTS: Thyroid-associated ophthalmopathy patients showed significant ocular surface damage (Rose Bengal staining score: TAO: 2.0 [0-5] versus controls: 0 [0-0.4]; IPC score: TAO 3.0 [0-8] versus controls: 0 [0-1], and BUT: TAO: 3.0 seconds [0-9 seconds] versus controls: 19.5 seconds [13-35.4 seconds]), and significantly reduced tear secretion (TAO: 10 mm [3-20 mm] versus controls: 17 mm [10-27 mm]). Ocular surface damage correlated significantly with tear secretion (r = - 0.35) but not significantly with mechanical alterations (impaired upgaze [r = - 0.34] and ocular surface increase [r = 0.32]). We firstly demonstrate that lacrimal acinar cells physiologically express TSHR. CONCLUSIONS: As ocular surface damage in TAO significantly correlates with reduced tear production, LG impairment appears to be a major cause of ocular surface drying. Intriguingly, physiological expression of TSHR by LG suggests that, in thyroid disease, autoantibodies may bind to lacrimal TSHR and, perhaps via aberrant signal transduction, contribute to LG impairment and, hence, dry eye syndrome.

Adult↗