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Ocular surface changes and discomfort in patients with meibomian gland dysfunction.

OBJECTIVE: To determine the importance of meibomian gland dysfunction (MGD) on the ocular surface. DESIGN: Prospective study. SETTING: A university-based referral practice. PATIENTS: Patients with ocular discomfort (147 eyes) and without ocular discomfort (54 eyes) were examined. In the total 201 eyes, MGD was defined as the presence of an obstruction of the meibomian orifices (obstruction group [n = 54]) or the absence of a gland structure (gland dropout group [n = 36]), or both of these findings (combined group [n = 38]). There were not any findings of MGD in 73 eyes (non-MGD group). MAIN OUTCOME MEASURES: Scores that were obtained from fluorescein and rose bengal staining, the breakup time of the tear film, the rates of tear evaporation and tear production, and meibography. RESULTS: Of the 147 eyes with ocular discomfort, 95 (64.6%) had either an obstruction of an orifice or gland dropout, or both. The combined group had higher scores for staining with fluorescein (P = .002) and rose bengal (P = .021) compared with that in the non-MGD group. The rate of tear production was increased more in the gland dropout group than in the non-MGD group (P = .002). The rate of tear evaporation was significantly increased in the gland dropout group (P = .017). CONCLUSION: Meibomian gland dysfunction is a major cause of ocular surface abnormalities and ocular discomfort.

Adult↗

Comparative study of 99mTc labeled hepatobiliary agents based on naphthalene and similar ring systems.

A series of 99mTc complexes of naphthalene and naphthalene-like ring systems were evaluated pharmacokinetically as hepatobiliary imaging agents and compared to 131I-rose bengal and 99mTc-N-(2,6-diethylacetanilide)-iminodiacetate in rats and dogs. Although all complexes studied exhibited high specificity for biliary excretion, they varied widely in disappearance rate from blood and in biliary excretion. Complexes with 8-hydroxyquinoline-7-carboxylic acid and hydralazine were relatively lipophilic and were eliminated more slowly than 131I-rose bengal; complexes of iminodiacetic derivatives of naphthalene, N-(2-acetylnaphthyl)iminodiacetic acid N-(2-naphthylmethyl)iminodiacetic acid were more polar and were eliminated much faster. Renal excretion of the complexes was 5.6% or less except for that of the latter compound (18%). Plasma binding ranged from 51 to 98%.

Animals↗

Treatment of experimental preretinal neovascularization using photodynamic thrombosis.

Retinal or preretinal neovascularization (NV) is the result of many ischemic conditions of the retina and is an important factor leading to severe visual loss in diabetic retinopathy. Panretinal photocoagulation does not always control its growth or bleeding sequelae. A potential new treatment modality, photodynamic therapy (PDT), was evaluated for limiting the progression of experimental NV in the rabbit eye. The NV was produced by injecting cultured dermal fibroblasts into the preretinal vitreous space after combined enzymatic and mechanical vitreolysis. This method results in traction retinal detachment with a rapid and consistent growth of NV. After administration of the photosensitizing dye rose bengal (20 mg/kg intravenously), PDT was done using a slit-lamp light source focused through a fundus contact lens (45 J/cm2). The NV was treated on two separate occasions during the active phase of growth (on days 13 and 21 after fibroblast injection). Control animals were exposed to light before injection of rose bengal. Eight randomly assigned animals in each group were followed between treatments and for 28 days after the second treatment. The appearance of NV was documented by frequent photography and fluorescein angiography. The PDT resulted in thrombosis of NV for at least 3 days. Reperfusion, however, was consistently noted at 7 days. Thrombosis was associated with a delay in the growth and maturation of NV fronds, which resumed after reperfusion. Twenty-eight days after the second treatment, NV in both experimental and control eyes had undergone atrophy. At that time (the conclusion of follow-up), however, the size of treated NV fronds (estimated from computerized image analysis of fluorescein angiograms) was significantly less than that of controls.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Interaction of singlet oxygen with 5'-nucleotidase in rat hearts.

This study was aimed to determine whether singlet oxygen (1O2) attenuates 5'-nucleotidase activity in the ischemic myocardium. Isolated rat hearts were exposed to either exogenous 1O2 produced by irradiating rose bengal or 40-min ischemia and reperfusion. Ecto-5'-nucleotidase activity was inhibited by exogenous 1O2 (3.74 +/- 0.38 mumol/min/g dry weight), when compared with normal control (7.52 +/- 0.41 mumol/min/g dry weight; P < 0.05). The enzymatic activity was significantly preserved by histidine (25 mM)--a 1O2 scavenger (7.04 +/- 0.61 mumol/min/g dry weight; P < 0.05 v rose bengal group). After ischemia, the activity of ecto-5'-nucleotidase was greatly reduced (2.51 +/- 0.25 mumol/min/g dry weight), when compared with normal control. Histidine significantly enhanced ecto-5'-nucleotidase activity (6.55 +/- 0.52 mumol/min/g dry weight, P < 0.05 v ischemic control). Adenosine release was consistent with ecto-5'-nucleotidase activity. The time course studies of effects of 1O2 on coronary flow, cardiac function, and LDH release revealed that the damage by 1O2 to ecto-5'-nucleotidase activity and adenosine release primarily accounted for impaired coronary flow, cardiac dysfunction, and impaired cardiac metabolism. Lipid peroxidation induced by exogenous 1O2 or ischemia was in parallel with ecto-5'-nucleotidase deactivation by 1O2. It is concluded that 1O2 causes inactivation of ecto-5'-nucleotidase and attenuation of adenosine release which could possibly be one of the important mechanisms of oxygen radical-mediated myocardial injury.

5'-Nucleotidase↗

The repeatability of clinical measurements of dry eye.

PURPOSE: The repeatability of individual dry eye diagnostic tests has been reported in the literature on normal samples of patients and to a lesser degree in dry eye patients. In this study, the repeatability of a battery of clinical diagnostic tests for dry eye was assessed on mild to moderate dry eye patients. METHODS: A dry eye examination was performed on 75 patients on two occasions by a single examiner. The battery of dry eye tests included symptom assessment, contact lens and medical history, slit-lamp biomicroscopic evaluation of the eyelids, evaluation of Meibomian glands, assessment of tear film quality, tear meniscus height, assessment of blink quality, fluorescein tear breakup time (TBUT), fluorescein and rose bengal staining of the cornea and conjunctiva, phenol red thread test, and Schirmer test. RESULTS: The repeatability of subjective report of dryness (kappaW = 0.62) and grittiness (kappaW = 0.73) was moderate to high. In contrast, the repeatability of Meibomian gland disease classification (kappaW = 0.20), presence or absence of inferior corneal fluorescein staining (kappa = 0.25), and inferior conjunctival rose bengal staining (kappa = 0.21) was poor. When a summed staining score of corneal and conjunctival regions was evaluated, weighted kappa reliability was fair. The repeatability of tear breakup time was substantial [95% limits of agreement -5.71 to 5.83 seconds; intraclass coefficient coefficient (ICC) 0.65], and improved when the two timed readings were averaged. Repeatability of the Schirmer test is more variable as wetting scores increase. When the average Schirmer scores </= 10 mm were evaluated, moderate repeatability was demonstrated, indicating that the test performs better with more advanced disease. CONCLUSION: Although patient-reported symptoms are moderately repeatable from visit to visit, many of the procedures clinically used to diagnose and monitor dry eye syndromes are largely unrepeatable.

Adult↗

Insecticidal activity of photoactive dyes to American and migratory grasshoppers (Orthoptera: Acrididae).

Many photoactive dyes are relatively nontoxic to vertebrates despite their insecticidal properties. Several photoactive dyes known to be toxic to some groups of insects were evaluated at various concentrations for toxicity to American and migratory grasshoppers in laboratory and field studies. Rose bengal and phloxine B were effective at inducing mortality of grasshoppers when applied at 2 and 5% to bran bait, though erythrosin B and uranine were ineffective. Partial replacement of phloxine with uranine in dye mixtures resulted in no significant loss of efficacy. Some indication of feeding inhibition was observed at high dye concentrations, so minimum effective dosages, probably 2%, are optimal. Phloxine B and rose bengal appeared to be stable upon exposure to sunlight, and able to withstand at least 24 h of sunlight without significant degradation. Dyes such as phloxine B could be a viable grasshopper control agent for small or medium-sized grasshopper species because mortality can be induced by consumption of a single flake dusted with 5% dye, and yet pose little hazard to vertebrates. Large species such as American grasshopper must consume several flakes before mortality is induced.

Animals↗

Silicone versus collagen plugs for treating dry eye: results of a prospective randomized trial including lacrimal scintigraphy.

PURPOSE: To compare the short-term efficacy of collagen and silicone plugs for treating dry eye using quantitative lacrimal scintigraphy. DESIGN: Prospective randomized clinical trial. METHODS: In this institutional study, 24 dry eye patients were evaluated in two groups: group I (n = 22 eyes) received collagen plugs and group II (n = 26 eyes) received silicone plugs. Comparisons were made with normal control subjects (n = 22 eyes). Data for the Schirmer I test, tear break-up time, and ocular surface staining with rose bengal dye were recorded before and after punctal occlusion. Lacrimal scintigraphy was performed at each time point, and the time to half maximum activity on the ocular surface (T(1/2)), and the percentage retention of activity on the ocular surface at the end of the dynamic study (RI) were recorded. RESULTS: In both patient groups, Schirmer I results, tear break-up times, and rose bengal staining scores improved significantly after plug insertion. Mean T(1/2) values and RI values increased significantly in both groups (P < .0001 for both). The differences for these values between groups I and II was statistically insignificant (P > .05). CONCLUSIONS: Collagen and silicone plugs both resulted in significant increases in aqueous tear volume, half-life of nuclear material on the ocular surface, and percentage of nuclear material retention. The groups' post-insertion values for all parameters were similar. These results suggest that these two plug types have similar efficacy as treatments for dry eye in the short term. Further studies evaluating long-term results are required.

Absorbable Implants↗

Glutathione S-transferases YcYfetus and YcYc--kinetic and inhibitor studies relating to their glutathione peroxidase activities.

The kinetic properties of the glutathione (GSH) peroxidase activities of GSH S-transferases YcYfetus and YcYc were compared. The catalytic efficiency of the fetal iso-enzyme with cumene hydroperoxide as substrate was approximately four times higher than the other. The effects of the non-substrate ligand rose-Bengal as well as the substrate ligands sulphobromophthalein and acrolein on the GSH peroxidase activity of these two iso-enzymes were also investigated. Depending on the ligand, the inhibition profiles of these two iso-enzymes when measured with either the peroxidase substrate, cumene hydroperoxide or the standard GSH S-transferase substrate 1-chloro-2,4-dinitrobenzene were found to be either very similar (sulphobromophthalein) or markedly different (rose Bengal and acrolein). Significantly, the GSH peroxidase activity of the fetal iso-enzyme was far less susceptible to inhibition by the teratogen, acrolein, than that of the YcYc isoenzyme. It is therefore attractive to suggest that should a similar situation arise in vivo, this resistance to peroxidase inhibition may play a role in preventing the fetotoxic effects of acrolein.

Acrolein↗

Tear film instability induced by rigid contact lenses.

PURPOSE: To evaluate rigid contact lens-associated epithelial damage, the relationship between the tear film and the ocular surface epithelium was evaluated at the perilimbal area around rigid contact lenses (RCLs). METHODS: Twenty RCL [six polymethylmethacrylate (PMMA) and 14 rigid gas-permeable lens (RGPL)] wearers and 20 non-CL wearers were enrolled in this study. By using a newly developed tear interferometer, the noninvasive tear film break-up time (NIBUT) on the conjunctiva at the 3- and 9-o'clock portions was measured, and its stability evaluated. Fluorescein and sulforhodamine B staining was used to observe ocular-surface epithelial damage, and rose bengal staining was used to evaluate the mucin layer distribution. RESULTS: The conjunctival NIBUT at the 3- or 9-o'clock portions or both was abnormally shortened in 15 (75%) of 20 RCL-wearer eyes. Conjunctival epithelial staining at the 3- or 9-o'clock portion was seen in six (30%) of 20 eyes by using sulforhodamine B, and in four (20%) of 20 eyes by rose bengal; all of these eyes also demonstrated an abnormal tear film breakup. Corneal epithelial damage was seen in two (10%) of 20 RCL-wearer eyes at the 3- or 9-o'clock portions or both, and in both of these cases, conjunctival epithelial damage also was noted. Neither tear-film instability nor epithelial staining was seen in the normal controls. CONCLUSIONS: This study indicates that RCL-induced tear-film instability is associated with damage to the ocular-surface epithelium and mucin layer.

Adult↗

Tear function and ocular surface changes with topical mitomycin (MMC) treatment for primary corneal intraepithelial neoplasia.

PURPOSE: To evaluate the tear function and ocular surface alterations in patients with primary CIN before and after treatment with topical mitomycin (MMC). PATIENTS AND METHODS: We describe seven patients with unilateral CIN treated with 0.04% topical MMC three times daily until full eradication of the lesion. The patients underwent tear and ocular surface examinations including Cochet-Bonnet corneal sensitivity measurements, tear film break-up time (BUT), Schirmer test, and Rose-Bengal staining before, at the time of resolution of the CIN, and at the final follow-up. Conjunctival impression cytology was performed before treatment and at the last visit. RESULTS: The mean pretreatment corneal sensitivity was 30.3 +/- 7.4 mm and improved to 55 +/- 5 mm at the final visit (P < 0.05). There were no aqueous-deficient eyes. The BUT values and Rose-Bengal staining scores also showed significant improvement at the last follow-up compared with the pretreatment values (P < 0.05). Initial impression cytology specimens showed goblet cell loss, higher grades of squamous metaplasia, areas of isolated keratinized, binucleated, and actively mitotic disfigured epithelial cells in all patients. The mean goblet cell density and squamous metaplasia grade were observed to improve significantly at the last visit (P < 0.05). MMC-induced cytologic changes were seen to persist long after cessation of treatment in some patients. All eyes remained free of recurrence and complications during the follow-up period. CONCLUSION: We found 0.04% topical MMC treatment tid until full eradication to be effective in the management of CIN. The ocular surface disease of CIN was characterized by disturbance of tear film stability, goblet cell loss, and increased squamous metaplasia in all patients. Impression cytology proved useful in attaining the diagnosis of CIN, evaluating the effect of treatment, and showing MMC-related long-term changes on the ocular surface.

Administration, Topical↗

The relation between tear film tests in patients with dry eye disease.

PURPOSE: The purpose of this report was to investigate the relation between dry eye diagnostic tests. METHODS: Dry eye patients were enrolled to complete a clinical examination, including the following dry eye tests: a meibomian gland evaluation, tear meniscus height, fluorescein tear breakup time, fluorescein staining of the cornea, the Schirmer 1 test, the phenol red thread test, and rose bengal staining of the conjunctiva. Statistical analyses, including correlation coefficients, the Wilcoxon sign rank test, chi-square test, and logistic regression were used to address the relation between these clinical tests of dry eye. RESULTS: There was a strong relation between the Schirmer test and fluorescein staining in all four statistical analyses. Similarly, there was also a strong relation between the phenol red thread test and both fluorescein and rose bengal staining. Finally, the results of the Schirmer test were associated with the tear breakup time test in three of four analyses. CONCLUSIONS: The results indicate that tests of aqueous deficiency (volume or production) are associated with ocular surface desiccation. This important relation should be recognized when choosing dry eye tests as outcomes in clinical trials and epidemiological studies.

Coloring Agents↗

A comparison of the sensitivity of photodamage assays in rat basophilic leukemia cells.

Many aspects of cellular function or physiology can be used to indicate the level of damage resulting from the application of potentially deleterious agents such as drugs, solvents or even light. The dose required to reach a specific biological endpoint will necessarily depend on the characteristics of the damage induced by the agent. By using multiple biological probes, it is possible to get a more complete description of the type of damage induced. Photodamage was induced in rat basophilic leukemia cells by either 254-nm UVC light exposure or rose bengal photosensitization. Damage was measured by three quantitative assays employing fluorescent probes: calcein, to measure nonspecific esterase activity, propidium iodide (PI), to measure loss of plasma membrane integrity, rhodamine 123 (R123) to measure mitochondrial depolarization, and the incorporation of 5'-bromodeoxyuridine (BrdU), to measure the progress of cell replication. BrdU incorporation was found to be the most sensitive indicator for both forms of photodamage. For UVC photodamage, the BrdU assay was 330 times more sensitive than the other two assays. For rose bengal photosensitization, the BrdU assay was 48 or 62 times more sensitive than either the R123 or calcein/PI assays, respectively.

Animals↗

The sensitivity and specificity of Brucella agglutination tests.

Brucellosis is a systemic infectious disease caused by Gram-negative bacilli, the genus Brucella, and clinical features are diverse. Therefore, several infectious and non-infectious diseases are considered in its differential diagnosis. In this study, we aimed to determine the positivity rate of Brucella agglutination tests in the culture-positive brucellosis and in diseases mimicking brucellosis clinically.Thirty patients with culture-positive brucellosis, and 280 patients with the diseases mimicking brucellosis clinically (20 with miliary tuberculosis, 33 with malaria, 20 with typhoid fever, 20 with adult-onset Still's disease, 47 with systemic lupus erythematosus, 50 with rheumatoid arthritis, 27 with sarcoidosis, and 63 with active lymphoma) were included in the study. Brucella agglutination tests (Rose-Bengal and Wright) were studied in serum samples of these 310 patients. Both Rose-Bengal and Wright tests (the latter in a titer of 1/160 or higher) were positive in all patients with brucellosis. For the other diseases, the test was slightly positive (1/40) in one patient with malaria and another with non-Hodgkin's lymphoma, and weakly positive (1/20) in a patient with typhoid fever. It remained negative in the remaining. In conclusion, agglutination tests currently used in the diagnosis of brucellosis are very sensitive and specific. Brucellosis can be effectively excluded from the diseases having similar clinical features by the use of agglutination tests.

Agglutination Tests↗

Arterial thrombosis model with photochemical reaction in guinea-pig and its property.

We have already developed an arterial thrombosis model in the rat femoral artery which utilized photochemical reaction between systemically injected rose bengal and transillumination of a green light with 540 nm wave length from the outside of the vessel. In the present study, we applied this model to guinea-pigs in order to produce a more suitable thrombus model for evaluation of antithrombotic drugs which act on the prostaglandin cascade. In the guinea-pigs, the irradiated femoral artery was completely occluded in 7 min after the injection of rose bengal (10 mg/kg) in a similar manner to the rats. The processes of primary endothelial injury and the subsequent formation of thrombus during this manipulation were observed by the electron microscopy. Pretreatment with aspirin and Y-20811, a thromboxane synthetase inhibitor, significantly prolonged the time required for occlusion in the guinea-pigs, while these drugs were ineffective in the rats. The antithrombotic effect of vapiprost, a thromboxane A2 receptor antagonist, was more pronounced in the guinea-pigs than the rats. In conclusion, this model in guinea-pigs is more suitable for evaluating antithrombotic drugs, particularly, the action of which is exerted involving the prostaglandin cascade.

Animals↗

The comparison of efficacies of topical corticosteroids and nonsteroidal anti-inflammatory drops on dry eye patients: a clinical and immunocytochemical study.

PURPOSE: To investigate whether conjunctival inflammation represents a primary event in the pathogenesis of keratoconjunctivitis sicca or whether it is a secondary inflammatory reaction caused by enhanced mechanical irritation as a result of surface dryness and whether anti-inflammatory drops (corticosteroids and nonsteroidal anti-inflammatory) have therapeutic effects and are similar. DESIGN: Single-masked, randomized, prospective clinical trial. METHODS: Thirty-two keratoconjuctivitis patients with or without Sjögren syndrome were included in the study. The patients were randomized to three groups. Group 1 patients received a topical artificial tear substitute (ATS); group 2 received ATS plus nonsteroidal anti-inflammatory drops (NSAID); and group 3 received ATS plus topical corticosteroidal drops. The eye symptom severity scores, Schirmer test values, rose bengal and fluorescein staining scores were evaluated before treatment and 15 and 30 days after start of treatment. Impression cytology specimens were stained using immunohistochemical methods to detect the percentages of human leukocyte antigen II (HLA-DR) positive, Apo 2.7 positive, and periodic acid-Schiff positive cells. Statistical analyses were performed within and between groups. Group 3 patients had significantly lower symptom severity scores, fluorescein and rose bengal staining, and HLA-DR positive cells on days 15 and 30 compared with patients in other groups. They also had a significantly higher number of periodic acid-Schiff positive (goblet) cells in their impression cytology specimens on days 15 and 30 compared with the other patients. On day 30, group 3 patients had significant differences compared with their baseline measurements in terms of above-mentioned parameters. However, we did not detect a significant effect of any treatment schedule on the Shirmer test value and the numbers of Apo 2.7 cells in impression cytology specimens. CONCLUSION: Topical corticosteroids had a clearly beneficial effect both on the subjective and objective clinical parameters of moderate-to-severe dry eye patients. These effects were associated with the reduction of inflammation markers of conjunctival epithelial cells.

Administration, Topical↗

Evaluation of firefly luciferase bioluminescence mediated photodynamic toxicity in cancer cells.

PURPOSE: This work investigated whether fLuc-catalyzed oxidation of D-luciferin generates sufficient light to induce photodynamic toxicity in cancer cells. PROCEDURES: Light emission was assessed via cooled CCD (charge-coupled device) camera. Parental and fLuc expressing cancer cells were exposed to subtoxic concentrations of photosensitizers (Rose Bengal or hypericin) and D-luciferin, sunlight, or lamplight. Toxicity was assessed by MTT assay. RESULTS: fLuc expressing cells emitted up to 500-fold higher levels of photons than parental cell lines. Although exposure to photosensitizer and sunlight reduced survival of various cell lines, survival of fLuc expressing cells incubated with photosensitizer and D-luciferin, or photosensitizer and lamplight, did not differ significantly from parental or untreated cells. CONCLUSIONS: Contesting recent reports, fLuc bioluminescence does not generate sufficient photons to induce Rose Bengal or hypericin photodynamic toxicity in a range of malignant and nonmalignant cell lines, and is not suitable as a generalizable approach to antineoplastic therapy.

Animals↗

Photodynamic activation of ion transport through lipid membranes and its correlation with an increased dielectric constant of the membrane.

Illumination of biological membranes with visible light in the presence of membrane-active sensitizers (e.g. rose bengal) is known to inactivate transport proteins such as ion channels and ion pumps. In some cases, however, illumination gives rise to an activation of transport. This is shown here for ion channels formed by alamethicin in lipid membranes, and for porin channels, which were isolated from the outer membrane of E. coli (OmpC) and from the outer membrane of mitochondria (VDAC) and were reconstituted in lipid membranes. An activation (in the form of an increased conductance) was also observed in the presence of the cation carriers valinomycin and nonactin. The activation phenomena were only present, if the membranes were made from lipids containing unsaturated double bonds. Activation was reduced in the presence of the antioxidant vitamin E. We suggest that the activation of the different transport systems has a common physical basis, namely an increase of the dielectric constant, epsilon(m), of the membrane interior by the presence of polar oxidation products of photodynamically induced lipid peroxidation. Experimental evidence for an enhanced dielectric constant was obtained from the finding of a light-induced increase of the membrane capacitance in the presence of rose bengal.

Electrochemistry↗

Iodide iontophoresis as a treatment for dry eye syndrome.

BACKGROUND/AIMS: Among the causes related to the development or perpetuation and aggravation of dry eye disease, oxidative reactions may have a role in the pathogenesis of this disorder. Antioxidants, such as iodide, have shown a strong effect in preventing the oxidative damage to constituents of the anterior part of the eye. In this clinical trial the effectiveness of iodide iontophoresis and iodide application without current in moderate to severe dry eye patients was compared. METHODS: 16 patients were treated with iodide iontophoresis and 12 patients with iodide application without current for 10 days. Subjective improvement, frequency of artificial tear application, tear function parameters (break up time, Schirmer test without local anaesthesia), vital staining (fluorescein and rose bengal staining) as well as impression cytology of the bulbar conjunctiva were evaluated before treatment, 1 week, 1 month, and 3 months after treatment. RESULTS: A reduction in subjective symptoms, frequency of artificial tear substitute application, and an improvement in certain tear film and ocular surface factors could be observed in both groups. A stronger positive influence was seen after application of iodide with current (iontophoresis), as observed in a distinct improvement in break up time, fluorescein and rose bengal staining, and in a longer duration of this effect compared with the non-current group. No significant change in Schirmer test results and impression cytology were observed in both groups. CONCLUSIONS: Iodide iontophoresis has been demonstrated to be a safe and well tolerated method of improving subjective and objective dry eye factors in patients with ocular surface disease.

Adult↗