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PubMed2000

Frequency of dry eye diagnostic test procedures used in various modes of ophthalmic practice.

PURPOSE: There are many suggested dry eye diagnostic test batteries in the literature. However, clinicians use a wide variety of dry eye diagnostic tests in clinical practice due to a number of factors, including time constraints. In addition, there has been no systematic description of the standard of care in diagnosing dry eye in the literature. The purpose of this study is to determine the type and frequency of dry eye diagnostic tests used in various modes of ophthalmic practice across a spectrum of dry eye severity. METHODS: A total of 467 patient charts (patients with a previous dry eye diagnosis) were reviewed retrospectively to determine diagnostic test frequency. In reviewing patient charts, the following tests were identified as performed or not performed: symptom assessment, fluorescein staining, tear break-up time (TBUT), Schirmer test, rose bengal staining, and "tear assessment" (including quantity and quality of the tear meniscus). RESULTS: Dry eye diagnostic tests were used in the following frequencies across all modes of practice: symptom assessment (82.8%), fluorescein staining (55.5%), TBUT (40.7%), tear assessment (22.2%), Schirmer test (8.5%), and rose bengal staining (4.9%). When the clinics are pooled, two test procedures were used with the highest frequency (38.9%). The most commonly performed two-test procedure combination was a symptom assessment combined with fluorescein staining of the cornea (43.7%). CONCLUSIONS: Symptom assessment plays a large role in the diagnosis of dry eye. The current clinical standard of care for dry eye diagnosis includes the performance of at least one test procedure in addition to a symptom assessment.

Contrast Media↗
PubMed1985

Substrate and phenobarbital induction of the biliary excretion of exogenous organic anions in rats.

Phenobarbital pretreatment significantly increased the biliary excretion of both non-metabolizable organic anions (rose bengal, eosin and amaranth) and the metabolizable bromsulphthalein (BSP). Phenobarbital stimulated the biliary excretion of BSP due to enhanced conjugation of BSP with glutathione and increased biliary excretion of bromsulphthalein-glutathione conjugate (BSP-GSH). On the other hand, pretreatment with the non-metabolizable substrates (rose bengal, eosin and amaranth) failed to stimulate the biliary excretion rate of these organic anions. Pretreatment with BSP enhanced the biliary excretion of total BSP due to a stimulation of GSH-S-transferase activity which conjugates BSP with glutathione.

Amaranth Dye↗
PubMed1992

Photodynamic inactivation of an ion channel: gramicidin A.

The ion channel formed by the peptide gramicidin A in planar lipid membranes is inactivated by visible light in the presence of the photosensitizer Rose Bengal. This is concluded from the strong decrease of the membrane conductance by more than two orders of magnitude. Experiments performed at different oxygen concentrations, in the presence of the singlet oxygen quenchers beta-carotene or alpha-tocopherol indicate, that presumably a type I process between the dye Rose Bengal and the tryptophan residues of the gramicidin channel with a subsequent oxidation of the tryptophans is responsible for the loss of the conductance properties of the channel.

Amino Acid Sequence↗
PubMed2001

Laser in situ keratomileusis-induced neurotrophic epitheliopathy.

PURPOSE: To report two cases of laser in situ keratomileusis-induced neurotrophic epitheliopathy with punctate epitheliopathy and rose bengal staining of the corneal flap. METHODS: Interventional case reports. RESULTS: A 42-year-old woman and a 37-year-old man with no preoperative symptoms or signs of dry eye developed dry eye symptoms and bilateral punctate epithelial erosions as well as rose bengal staining of the corneal flaps after laser in situ keratomileusis. Neither patient had less than 12 mm of wetting with the Schirmer test without anesthesia at any time point between development and resolution of the flap surface abnormalities. The flap surface abnormalities resolved approximately 6 months after laser in situ keratomileusis. CONCLUSIONS: Laser in situ keratomileusis-induced neurotrophic epitheliopathy may be attributable to loss of trophic influence from severed corneal nerve trunks. The condition typically resolves approximately 6 months after laser in situ keratomileusis or laser in situ keratomileusis retreatment.

Adult↗
PubMed1994

MRI monitoring of experimental cerebral ischaemia: comparison of two models.

In experimental stroke research one of the key questions is whether animals models truly reflect human disease. One important reason for modelling stroke is to produce tissue changes that can be used to validate the correlation between the results of advanced radiological imaging and pathology. The aim of this study was to compare two minimally invasive rat models of focal ischaemic stroke as to their ability to simulate clinical stroke reproducibly. In one model a focal infarct was induced using the photochemical Rose Bengal technique. The second model was based on transvascular occlusion of the middle cerebral artery using a cervical approach. In all animals we performed MRI at different times with two different paramagnetic contrast agents. The time course of blood-brain-barrier disruption in the Rose Bengal model differed entirely from that in human brain ischaemia. The experimental stroke showed marked contrast enhancement in the first hour after the onset of ischaemia. On the other hand, the MRI changes in the suture occlusion model were very similar to the changes observed in human brain ischaemia: no early disruption of the blood-brain-barrier and increased T2-signal 4-6 h after the onset of stroke. We hope that the suture occlusion model will be used for research not only by basic and clinical scientists but also by radiologists, who, using modern imaging methods, may greatly influence the diagnosis as well as the management of this devastating disease.

Animals↗
PubMed1996

Does paracellular permeability play a role in cholephilic dye-induced cholestasis?

Changes in hepatic paracellular permeability were investigated during the development of cholephilic dye-induced cholestasis in rats. For this purpose, four dyes with different cholestatic potency (phenol red, sulfobromophthalein, bromcresol green and rose bengal) were infused at a high, potentially damaging dose (280 nmol/min per 100 g body wt., i.v.), and changes in paracellular permeability were continuously monitored by measuring the access into bile of the permeability probe -14C-sucrose. The cholestatic potency of the different dyes was: rose bengal > bromcresol green > sulfobromophthalein > phenol red. All dyes increased [14C]sucrose bile-to-plasma ratio, producing a displacement towards curves of higher permeability. The capability of the dyes to increase biliary permeability followed the same order as their respective cholestatic potencies. The possible implications of the present results for cholephilic dye-induced cholestasis are discussed.

Animals↗
PubMed1977

Internal and external application of photodynamic sensitizers on squid giant axons.

Squid giant axons were photosensitized by dyes applied internally or externally in air saturated solutions and photochemically modified by visible light. For most dyes the modifications included an irreversible block of sodium channels, a destruction of inactivation in some of the unblocked channels, and a slowing of inactivation. Internal application was up to 100-fold more effective in blocking sodium channels than external application, suggesting a site of block nearer the internal surface. Rose Bengal sensitized channel block and destruction of inactivation when applied internally, but sensitized only channel block when applied externally. In contrast, externally applied Eosin Y sensitized a clear slowing of inactivation plus channel block. Beta-carotene, an effective agent for quenching photochemically generated excited singlet oxygen, inhibited most of the modification sensitized by internally applied Methylene blue but not by Rose Bengal or Merocyanine 540.

Action Potentials↗
PubMed1985

Role of hydrogen peroxide in riboflavin-sensitized photodynamic damage to cultured rat lenses.

Previous reports have differed on whether hydrogen peroxide or free radicals are the principal toxic species responsible for photodynamic damage to intact lenses cultured in medium with riboflavin, light and oxygen. To differentiate between hydrogen peroxide and free radicals, TC-199 medium containing 50 microM riboflavin was incubated in visible light prior to adding the lenses. After 4 hr the medium was placed in the dark, rat lenses, [3H]-choline and 86Rb were added, and the lenses were cultured for 4 hr. Damage to the lenses was evaluated by measuring the accumulation of choline and Rb. Lenses cultured in previously irradiated medium accumulated less choline and Rb than lenses in control medium, indicating that in the presence of light, riboflavin generates a stable factor which is toxic to lenses. The lens damage was similar to that caused by direct exposure of lenses to riboflavin and light for 4 hr. This toxicity was blocked by catalase, and was presumed to be caused by hydrogen peroxide. Because of their short lifetime in solution, photochemically generated free radicals could not have been present in the dark after the lenses were added. Rose bengal, a photosensitizer which is thought to form singlet oxygen, did not generate a stable toxic factor in the culture medium and damaged lenses only if they were present during the exposure to light. The hydrogen peroxide concentration in riboflavin-containing medium after 4 hr exposure to light was approximately 200 microM, whereas rose bengal generated less than 10 microM hydrogen peroxide.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphate↗
PubMed2001

Focal ischemia in the cerebral cortex has an effect on the neurohypophysis. I. Ultrastructural changes in capillary vessels of the neurohypophysis after focal ischemia of the cerebral cortex.

OBJECTIVES: In our investigations we have reported that photochemical reaction leading to brain ischemia can also be precipitated with visible light from a non-coherent light source. It was revealed that focal cerebral ischemia after photochemical reaction cause the alterations in the capillaries ultrastructure and perivascular spaces of the barrier-competent regions of the brain. The purpose of this study is to first characterize the ultrastructural morphological consequences of photochemically induced ischemia in the cerebral cortex on the capillaries of neurohypophysis as the barrier-free region of the brain. METHOD: We used a model of ischemic brain damage due to obliteration of microvessels following the photochemical reaction. Rats were treated with an intravenous injection of rose bengal and irradiated from a halogen lamp source through an intact cranium to precipitate microvascular damage. Material for electron microscopic studies were sampled from the neurohypophysis 1 and 4 days after irradiation (4 animals in each group) in experimental group and 1 and 4 days after a rose bengal injection in control group. RESULTS: Investigations in transmission electron microscopy revealed platelet aggregation on the endothelium preceded by its early ultrastructural damage. In the capillaries of the neurohypophysis, one and four days after irradiation, numerous microthrombi adhering to the damaged endothelium were present. The capillary vessels contained a continuous, rather than a fenestrated endothelium. The basement membrane was thickened, blurred and locally multiplicated. CONCLUSION: Our results show that experimentally-induced thrombosis of cortical microvessels leads to alterations in the capillaries of neurohypophysis.

Animals↗
PubMed2003

[Experimental retinal vein occlusion induced by photodynamic approach].

OBJECTIVE: To develop a new photochemical method of experimental retinal vein occlusion and investigate the morphologic and histologic changes in the retina. METHODS: After intravenous injection of rose Bengal, the vessels next to the disc of miniature pigs in the experiment group (n = 15) were exposed to an endo-illuminator for 15 minutes. As a control group, the vessels next to the disc of each pig were exposed to the endo-illuminator without rose Bengal injection. After complete vascular occlusion, the eyes were observed at following times: one hour, and 1, 3, 7, 14, 21, and 28 days. After the 28th day, the eyes were enucleated and prepared for light and electron microscope examination. RESULTS: Histopathologic features were consistent with changes of retinal vein occlusion and formation of retinal vein thrombi was reliable. CONCLUSIONS: Photodynamic method combined with endo-illuminator was a simple, reliable and definitive experimental technique to produce retinal vein occlusion.

Animals↗
PubMed2003

Alterations in conjunctival cytology and tear film dysfunction in patients with beta-thalassemia.

PURPOSE: Patients with beta-thalassemia (beta-tha) represent a group with lifelong transfusion-dependent anemias. This study aimed to describe the conjunctival changes and tear film parameters in these patients. METHODS: A total of 52 patients (104 eyes) with beta-tha major and 22 normal control subjects (44 eyes) were studied during 1999 through 2000. Tear film break-up time (BUT), Schirmer test, rose Bengal staining, and cytologic evaluation of the conjunctival epithelium were performed in all subjects. The Papanicolaou and May-Grümwald-Giemsa staining procedures were performed on all smears. Patients and control subjects were compared for tear function parameters and conjunctival changes. RESULTS: The BUT, Schirmer test, and rose Bengal staining values were significantly lower (P < 0.001) in beta-tha patients than in control subjects. Keratinized cells were observed in conjunctival samples in 41% of patients, with a decrease in the number of goblet cells per slide in 64% of patients. In 9% of beta-tha patients, there were a slightly greater number of inflammatory cells than in control eyes. CONCLUSION: Ocular surface disorder of these patients was characterized by goblet cell loss and conjunctival squamous metaplasia. Our findings were correlated positively with the variable age. Epithelial damage by toxic reaction and disorder of tear quality and quantity are implicated as important factors in the pathogenesis of the ocular surface disease in beta-tha patients.

Adolescent↗
PubMed2002

Destruction of fecal bacteria in wastewater by three photosensitizers.

Agriculture in Tunisia faces acute problems of water quality and quantity caused by limited conventional water resources. One possibility to cope with low water resources is to purify wastewater for reuse. Among different methods for the disinfection of wastewater, the inactivation of fecal coliforms using a combination of a photosensitiser (Rose Bengal, Methylene Blue, cationic porphyrin) with sunlight was determined on a small scale. In parallel the sensitizer photobleaching was also followed under the same conditions. The results described in this paper show that the meso-substituted cationic porphyrin is more efficient and more photostable than Methylene Blue and Rose Bengal in wastewater. A lower cationic porphyrin concentration, 1 microM, resulted in very little cell death. Higher concentrations, 5 microM or 10 microM, produced more cell death. Nevertheless there was a small difference between concentrations 5 and 10 microM. By increasing the duration of irradiation we can improve the log reduction in bacteria and compensate for a low concentration of sensitizer or for a less efficient type of sensitizer. The same log reduction in fecal bacteria was obtained with 5 and 10 microM of cationic porphyrin during the fourth hour of treatment.

Agriculture↗
PubMed1992

Singlet oxygen-induced inhibition of cardiac sarcolemmal Na+K(+)-ATPase.

We investigated the susceptibility of sarcolemmal Na+K(+)-ATPase to singlet oxygen. The role of this enzyme is regulation of Na+ concentration and thereby membrane potential. Inhibition of Na+ pump would lead to intracellular Ca2+ overload therefore further aggravating the injury caused by free radicals. Incubation of isolated sarcolemmal vesicles with irradiated rose bengal (150 nM) resulted in 86 +/- 1% inhibition of Na+K(+)-ATPase activity and histidine (25-100 mM) protected the enzyme in a dose-dependent fashion whereas SOD, catalase or mannitol (.OH radical scavenger) did not have any effect. Also, the inhibition of Na+K(+)-ATPase activity was dependent on rose bengal concentration, intensity of irradiation, duration of light exposure, showing that inhibition was directly related to amount of singlet oxygen generated. These results show that singlet oxygen may have significant disruptive effects on sarcolemmal function and may represent an important mechanism by which the oxidative injury to the myocardium induces arrhythmogenesis.

Animals↗
PubMed1984

Absorption of 5-methyltetrahydrofolate in rat jejunum with intact blood and lymphatic vessels.

UNLABELLED: Transport results from in vitro studies may not be applicable to in vivo situations. In this study, we extended our previous in vitro observations regarding the intestinal transport of 5-methyltetrahydrofolate to in vivo studies in the unanesthetized rat and examined the effect of the unstirred water layer on the absorption process. We used a well defined intestinal perfusion technique. Absorption of 0.5 and 5 microM 5-methyltetrahydrofolate proceeded in a linear manner for 40 min of perfusion at 0.31 and 1.74 nmol/100 cm per min, respectively. Absorption of 0.5 microM 5-methyltetrahydrofolate increased with increasing perfusate flow-rate from 0.5 to 2 to 4 ml/min, indicating an unstirred water layer influence on the absorption rate. Absorption of the substrate was saturable with an apparent Kt of 5.7 microM and Vmax of 3.45 nmol/100 cm per min. Absorption was pH-dependent, and was inhibited by structural analogues. In contrast to the in vitro data, addition of glucose (20 mM) to the perfusate was unnecessary for in vivo absorption to proceed. Unconjugated cholic (5 mM) and deoxycholic (1 mM) acids and the organic anion rose bengal (0.1 mM) inhibited the absorption of 0.5 microM 5-methyltetrahydrofolate when added to the perfusate. CONCLUSIONS: the results of previous in vitro studies of 5-methyltetrahydrofolate intestinal transport are applicable to in vivo situations, except that luminal glucose was found to be unnecessary in the latter. The unstirred water layer modulated the absorption of 5-methyltetrahydrofolate, while unconjugated bile acids and rose bengal inhibited it.

Animals↗
PubMed1997

Preretinal neovascularization induced by experimental retinal vein occlusion in albino rats.

Retinal ischemia and neovascularization have been demonstrated in several animal models. To determine 1) whether the retinal or preretinal neovascularization can be induced in albino rats by retinal vein occlusion and 2) the type and rate of occurrence on neovascularization, we occluded retinal veins in albino rats by photodynamic thrombosis. After anesthesia, each of 36 rats received an injection of rose bengal photosensitive dye, and their veins underwent argon green laser treatment. Half or all the major retinal veins were occluded in 12 eyes and in 24 eyes, respectively. Ten control rats underwent the same procedures but the laser beam was directed between major retinal vessels. In 46 control eyes, rose bengal dye was seen to have perfused without laser treatment. Retinal detachment developed in most vein occluded eyes within one day of venous occlusion, which was confirmed by fluorescein angiography. On follow-up at two weeks, only four of 24 eyes (16.7%) had undergone occlusion of all retinal veins showed new preretinal vessels on the optic disc. In these four eyes, severe disturbance of both retinal arterial and venous blood flow was observed, but no other eyes showed such severe combined disturbance. These data suggest that preretinal neovascularization in albino rats can be induced by this minimally traumatic method and that venous occlusion is severe enough to compromise arterial blood flow for a certain threshold period, thus inducing the development of preretinal neovascularization.

Animals↗
PubMed1989

Singlet molecular oxygen causes loss of biological activity in plasmid and bacteriophage DNA and induces single-strand breaks.

Damage of plasmid and bacteriophage DNA inflicted by singlet molecular oxygen (1O2) includes loss of the biological activity measured as transforming capacity in E. coli and single-strand break formation. Three different sources of 1O2 were employed: (i) photosensitization with Rose bengal immobilized on a glass plate physically separated from the solution; (ii) thermal decomposition of the water-soluble endoperoxide 3,3'-(1,4-naphthylidene) dipropionate (NDPO2); and (iii) microwave discharge. Loss of transforming activity was documented after exposing bacteriophage M13 DNA to 1O2 generated by photosensitization employing immobilized Rose bengal, and with bacteriophage luminal diameter X174 DNA, using the thermodissociable endoperoxide (NDPO2) as a source of 1O2. These findings are in agreement with experiments in which plasmid DNA pBR322 was exposed to a gas stream of 1O2 generated by microwave discharge. The effects of 1O2 quenchers and of 2H2O indicate 1O2 to be the species responsible. Strand-break formation in pBR322 and luminal diameter X174, measured as an increase of the open circular form at the expense of the closed circular supercoiled form, was observed without alkaline treatment after exposing the DNA to 1O2, using either agarose gel electrophoresis or sucrose gradient separation. The effect of quenchers and 2H2O indicate the involvement of 1O2 in DNA damage. We conclude that singlet oxygen can cause loss of biological activity and DNA strand breakage.

Azides↗
PubMed2004

Molecular aspects of plant adaptation to life in the Chernobyl zone.

With each passing year since the Chernobyl accident of 1986, more questions arise about the potential for organisms to adapt to radiation exposure. Often this is thought to be attributed to somatic and germline mutation rates in various organisms. We analyzed the adaptability of native Arabidopsis plants collected from areas with different levels of contamination around the Chernobyl nuclear power plant from 1986 to 1992. Notably, progeny of Chernobyl plants resisted higher concentrations of the mutagens Rose Bengal and methyl methane sulfonate. We analyzed the possible molecular mechanisms of their resistance to mutagens and found a more than 10-fold lower frequency of extrachromosomal homologous recombination, significant differences in the expression of radical scavenging (CAT1 and FSD3) and DNA-repair (RAD1 and RAD51-like) genes upon exposure to mutagens (Rose Bengal and x-rays), and a higher level of global genome methylation. This data suggests that adaptation to ionizing radiation is a complex process involving epigenetic regulation of gene expression and genome stabilization that improves plants' resistance to environmental mutagens.

Adaptation, Physiological↗
PubMed1982

Phylogeny and ontogeny of the phosphoglycerate mutases.--V. Inactivation of phosphoglycerate mutase isozymes by histidine-specific reagents.

1. The three isozymes of glycerate-2,3-P2 dependent phosphoglycerate mutase present in tissues of mammals and reptiles were inactivated by both treatment with diethylpyrocarbonate and photooxidation with rose bengal. 2. Inactivation of type M isozyme purified from rabbit muscle was complete when two histidine residues per enzyme subunit were carboethoxylated. Hydroxylamine removed the carboethoxy groups, with partial recovery of the enzymatic activity. The cofactor protected the enzyme against inactivation. 3. The inactivation of rabbit muscle phosphoglycerate mutase by photooxidation with methylene blue and rose bengal was sharply pH dependent. The pH profile of enzyme inactivation followed the titration curve of histidine, suggesting that this amino acid was critical for enzyme activity. Glycerate-2,3-P2 did not protect phosphoglycerate mutase against photoinactivation.

Animals↗