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PubMed1990

[Treatment of dry eye by temporary punctum plug. Comparison with a control group].

35 patients with chronic dry eye were treated with punctum plug on one side only. The other side was a control: after simulation dilatation, the patient underwent a simulation insertion. This is an open study in which only the patient ignored the reality of treatment. The study was prospective. 28 females and 7 males were enrolled in this study. The age' average was 53. The follow-up average is 5.3 months. The etiologies were: idiopathic dry eye: 10 cases, Sjögren's syndrome: 11 cases, drug induced dry eye: 6 cases, filamentous keratitis: 4 cases. Other: 4 cases. Enrollment criteria were: functional symptomatology not answering to topical therapy (physiological serum drops) associated with one or more of the following items: positive Rose Bengal staining and/or Schirmer lower than 5 mm in 5 min and/or filamentary keratitis. Subjective improvement is significantly better after closing the two puncta on the same side (77%) than the other side (placebo) (17%) (p less than 0.001). This difference is also demonstrated for the improvement of Rose Bengal staining (p = 0.068) and for Schirmer's test (p = 0.002). These results demonstrated the effectiveness of this therapy versus placebo. Results after closing the 2 puncta on the same side are better than after closing one punctum.

Adolescent↗
PubMed2003

[Long-term follow-up after autologous submandibular gland transplantation in scarring keratoconjunctivitis with absolute dry eyes].

INTRODUCTION: An autologous submandibular gland can be transplanted to the temporal fossa and following microvascular anastomosis to the temporal artery and vein and implantation of the secretory duct into the superotemporal conjunctival fornix to provide continuous substitute lubrication. Here we report our experiences with this procedure. PATIENTS AND METHODS: In a prospective, controlled clinical cohort study, we investigated the development of the Schirmer- test, fluorescein-break up time (F-BUT), degree of discomfort, use of pharmaceutical tear substitutes, visual acuity, Rose Bengal staining and conjunctival hyperemia in 14 eyes with a successful submandibular gland transplant and 11 dry eyes without salivary lubrication. RESULTS: Over a mean postoperative period of 3.3 years the transplantation group showed significant improvements of the Schirmer-Test, FBUT, use of pharmaceutical tear substitutes, discomfort and Rose Bengal staining up to the last follow-up when compared to the preoperative and control groups. CONCLUSION: Transplantation of an autologous submandibular gland can improve the lubrication and discomfort of absolute tear deficiency on a long term basis, but the salivary tear film only partially stabilises the ocular surface. The procedure is however able to provide symptomatic relief in extreme cases of absolute dry eyes.

Adolescent↗
PubMed2005

Adsorption of xanthene dyes by lysozyme crystals.

Adsorption characteristics of cross-linked lysozyme crystals of different morphologies (tetragonal, orthorhombic, triclinic and monoclinic) were examined using four anionic dyes (fluorescein, eosin, erythrosin, and rose bengal), one zwitterionic dye (rhodamine B), and one cationic dye (rhodamine 6G). The adsorption isotherms were of the Langmuir type for all examined systems with the exception of rhodamine B adsorption by monoclinic crystals. The weakest adsorption was observed for the cationic dye, rhodamine B, whereas dianionic dyes, eosin, rose bengal, and erythrosin were strongly adsorbed on the protein surface. The adsorption capacities of the crystals for the dyes were found to depend on both charge and hydrophobicity of the dye, reflecting the heterogeneous character of the lysozyme pore surface. The adsorption affinity of the crystals for the dyes was a function of the dyes' hydrophobicity. Furthermore, the crystal morphology was identified as an additional factor determining capacity and affinity for dye adsorption. Differences between crystals prepared in the presence of the same precipitant were lower than between morphologies prepared with different precipitants.

Adsorption↗
PubMed1999

Mitogen-activated protein kinase (p38-, JNK-, ERK-) activation pattern induced by extracellular and intracellular singlet oxygen and UVA.

Ultraviolet A (UVA; 320-400 nm) radiation in human skin fibroblasts induces a pattern of mitogen-activated protein kinase (MAPK) activation consisting of a rapid and transient induction of p38 and c-Jun-N-terminal kinase (JNK) activity but not extracellular signal-regulated kinases (ERK). UVA activation of p38 can be inhibited by the singlet oxygen (1O2) quenchers azide and imidazole, but not by the hydroxyl radical scavengers mannitol or dimethylsulfoxide, pointing to the involvement of 1O2. The same effect has been shown for JNK. Like UVA, 1O2 generated intracellularly upon photoexcitation of Rose Bengal activates p38 and JNK but not ERK. p38 and JNK activation was also elicited by chemiexcitation for the intracellular generation of 1O2 by the lipophilic 1,4-endoperoxide of N,N'-di(2,3-dihydroxypropyl)-1, 4-naphthalene dipropionamide. In contrast, extracellular generation of 1O2, by irradiation of Rose Bengal immobilized on agarose beads or by chemiexcitation employing the hydrophilic 1,4-endoperoxide of disodium 3,3'-(1,4-naphthylidene) dipropionate, was ineffective in activating p38 or JNK. These data suggest that the activation of p38 and JNK by 1O2 occurs only when the electronically excited molecule is generated intracellularly.

Calcium-Calmodulin-Dependent Protein Kinases↗
PubMedDate not supplied

Photokeratitis from subablative 193-nanometer excimer laser radiation.

BACKGROUND: Photokeratitis is a side effect of UV light exposure whereby the corneal epithelium is photochemically injured in a time delayed fashion. UV light exposure in the far ultraviolet wavelength range has not previously been observed. This study addresses the concern of photokeratitis from subablative 193-nanometer excimer laser light. METHODS: Dutch belted rabbit corneas were irradiated with subablative 193-nanometer excimer laser light over a wide range of total energy exposures, and examined by slit lamp biomicroscope for signs of photokeratitis. Photokeratitis was identified by epithelial haze and stippling, and rose bengal and fluorescein staining at varied time intervals between 1/2 to 26 hours post exposure. RESULTS: At threshold energy exposures of 1.0 to 1.5 J/cm2, an immediate superficial epithelial haze was seen which disappeared within several hours. At higher energy exposures of 10 J/cm2, a delayed photokeratitis with deep rose bengal and even fluorescein staining was seen. This latter delayed photokeratitis resembles that of longer UV wavelengths and is due to excimer laser fluorescence, whereas the former is a direct response of the 193-nanometer light. The percentage of excimer laser light undergoing fluorescence is calculated as less than 1%. CONCLUSIONS: The potential side effects and hazards of scattered 193-nanometer radiation during excimer laser surgery are extremely limited because of the shorter penetration depth of direct excimer radiation and the minimal fluorescent emission of longer UV wavelengths for energy exposures within the realm of clinical use.

Animals↗
PubMed1986

Photoinactivation of acetylcholinesterase by erythrosin B and related compounds.

Acetylcholinesterase was rapidly inactivated when exposed to light in the presence of xanthene dyes. Photosensitizing efficiency paralleled the dye triplet state quantum yields, increasing in the order fluorescein less than eosin B less than eosin Y less than erythrosin B less than rose bengal. The observed first-order rate constants of photoinactivation increased hyperbolically with dye concentration. Evidence for the formation of a dye-enzyme complex prior to inactivation was obtained from spectrophotometric and protein fluorescence quenching methods. The latter technique allowed estimates of the dye-enzyme dissociation constants for rose bengal (20 microM) and erythrosin B (30 microM). After photoinactivation, a portion of the dye became covalently bound to the enzyme. The photoinactivation reaction occurs in both aerobic (air saturated) and anaerobic (argon saturated) solution, with the rates of photoinactivation being about three to five times greater under the latter conditions. The aerobic reaction exhibits a large deuterium isotope enhancement effect and is largely (but not completely) quenched by 10(-2) M azide. The anaerobic reaction is unaffected by azide and exhibits only a small deuterium isotope effect. These results indicate that the photoinactivation reaction proceeds mainly by a type II (singlet oxygen mediated) pathway under aerobic conditions and by a type I (radical) pathway under anaerobic conditions. The enzyme was protected from inactivation by edrophonium, a competitive inhibitor, but not by d-tubocurarine, a peripheral-site ligand, indicating that destruction of a crucial residue at or near the catalytic site is an important component of the inactivation process. Extensive destruction of tryptophan undoubtedly occurs, at least under aerobic conditions.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcholinesterase↗
PubMed1995

[Diagnosis and and differentiation of dry eye disorders].

Artificial tears often fail in dry eye patients. Our goal was to establish a diagnostic approach that involves alternatives that are more effective for the treatment. We examined 110 patients with dry eyes not stabilized by artificial tears: medical history, visual acuity, slit lamp examination, rose bengal stains and fluorescein stains, Schirmer test, break-up time (BUT), dye tests, impression cytology, and lid transillumination. This allowed disturbances of the three layers of the tear film (lipid, aqueous, mucin) to be differentiated. Cellular damage of the ocular surface was evaluated and scored by slit lamp examination, rose bengal staining, and impression cytology. Only 8% of the patients with "sicca syndrome" had exclusive aqueous deficiencies amenable to artificial tears. In contrast, lipid deficiencies (chronic blepharitis) were determined most frequently (78%). Twenty-six percent had disturbances in two or more layers of the tear film. The diagnostic strategy of differentiating disturbances of the layers of the tear film offers a more specific approach to the treatment of dry eyes.

Adult↗
PubMed1993

Effects of organic anions and bile acids on biliary lipid excretion in hyperbilirubinemic mutant Sprague-Dawley rats.

The effects of organic anions and bile acids on biliary lipid excretion were studied in EHBR, a hyperbilirubinemic mutant Sprague-Dawley rat. A marked delay in the biliary excretion of BSP, cefpiramide, rose bengal and ursodeoxycholate-disulfate was observed in these animals. The marked decrease in the biliary excretion of phospholipids and cholesterol and the uncoupling of biliary bile acids and lipids that occurred after the administration of BSP, cefpiramide and ursodeoxycholate-disulfate in control Sprague-Dawley rats was absent in EHBR. Rose bengal did not change biliary lipid excretion in either the control Sprague-Dawley rats or the EHBR. Although taurocholate markedly increased bile flow and biliary bile acid excretion in both types of rats, the increase in biliary lipid excretion observed in the control Sprague-Dawley rats was absent in EHBR. These findings indicate that EHBR have an impairment of hepatic lipid transfer that is enhanced by bile acids, possibly at the level of intracellular vesicular lipid transport.

Animals↗
PubMed1987

Transport of conjugated bilirubin and other organic anions in bile: relation to biliary lipid structures.

Using gel-permeation chromatography, we studied associative relationships between conjugated bilirubin and various biliary lipid particle species, including lecithin/cholesterol vesicles, mixed-lipid micelles, and simple bile salt micelles. Five other organic anions were comparably studied: phenol red, Evans blue, sulfobromophthalein, rose bengal, and indocyanine green. For compounds of intermediate hydrophobicity, including conjugated bilirubin, the dominant association was with a bile salt/organic anion hybrid particle of dimensions larger than that of a simple pure bile salt micelle. Vesicular association was found to be dominant only for the most hydrophobic organic anions, indocyanine green and rose bengal; conversely, the most hydrophilic anion, phenol red, showed no vesicular association. Accordingly, a strong positive correlation (P less than 0.001) was found between percent vesicular association and degree of hydrophobicity of the organic anion. Alkaline conditions (eluant pH 9) decreased or prevented vesicular hydrophobic interaction with all anions. We conclude that two important particulate mechanisms for transport in bile of conjugated bilirubin and other water-soluble anions are bile salt/organic anion hybrid particles and vesicles. For most organic anions of intermediate hydrophobicity, including conjugated bilirubin, the bile salt/organic anion hybrid particle is the dominant transport vehicle.

Bile↗
PubMed1983

The prelaparotomy diagnosis of extrahepatic biliary atresia.

The diagnostic accuracy of laboratory investigations in the prelaparotomy differentiation between extrahepatic biliary atresia (EHBA) and intrahepatic disease (IHD) was assessed in 86 consecutive infants presenting with conjugated hyperbilirubinaemia. Forty five infants had EHBA and 41 IHD. The mean serum bilirubin concentration, gamma-glutamyltranspeptidase (GGT) activity, and the GGT/aspartate transaminase (AST) ratio were appreciably higher in infants with EHBA than in those with IHD. In infants with IHD, however, serum bilirubin concentrations were in the EHBA range in 19 (47%), as were GGT values in 29 (71%), and GGT/AST ratios in 33 (80%). In individual patients neither increasing nor decreasing GGT values were of diagnostic importance. Failure of biliary excretion of 99Tcm-p-Butyl-ida occurred in 29 of 30 (97%) patients with EHBA but also in 22 of 23 (67%) with IHD. In all 5 patients with IHD associated with alpha 1 antitrypsin deficiency these 4 investigations gave results in the EHBA range. Liver biopsy specimen interpretation, correct in 38 of 42 infants with EHBA, gave an overall accuracy of diagnosis of 86%: the results of 3 further biopsies were equivocal. In 33 of 40 infants with IHD bile duct obstruction was excluded; the remaining 7, including 4 with alpha 1 antitrypsin deficiency, showed equivocal changes. Faecal excretion of 131I rose bengal faecal excretion was less than 10% in 36 of 37 patients with EHBA and in 9 of 26 with IHD, giving an overall accuracy of diagnosis of 84%. In patients in whom genetic disorders, such as alpha 1 antitrypsin deficiency had been excluded, interpretation of liver biopsy specimens together with 131I rose bengal faecal excretion remain the most accurate means of identifying those who need surgery for EHBA and of avoiding unnecessary laparotomy in infants with IHD.

Aspartate Aminotransferases↗
PubMed1996

Simplified model of krypton laser-induced thrombotic distal middle cerebral artery occlusion in spontaneously hypertensive rats.

BACKGROUND AND PURPOSE: The effects of thrombotic occlusion of the middle cerebral artery on compromised ischemic tissue may be different from and more severe than those of cerebral ischemia induced by mechanical occlusion of the artery. Photothrombosis, which is based on photochemical damage to the endothelium and subsequent platelet aggregation, is an efficient method to induce thrombosis in vivo. This study aimed to improve and simplify this unique method for an ischemia model of middle cerebral artery occlusion in rats. METHODS: Male spontaneously hypertensive rats (5 to 6 months old, 300 to 450 g) were anesthetized with halothane, endotracheally intubated, and mechanically ventilated. A krypton laser operating at 568 nm was used to irradiate the exposed distal middle cerebral artery with an intact dura above the rhinal fissure. The photosensitizing dye rose bengal (20 mg/kg body wt) was administered intravenously over 90 seconds starting simultaneously with 4 minutes of laser irradiation at a power of 20 mW to cause thrombotic occlusion of this artery. RESULTS: The irradiated middle cerebral artery was completely occluded by intraluminal thrombi within 3 minutes after simultaneous laser irradiation and rose bengal infusion. Thrombosed materials were not stained by phosphotungstic acid-hematoxylin stain (ie, aggregated platelets lacked apparent fibrin). The mean volume of 3-day-old infarction, indicated by the lack of staining with 2,3,5-triphenyltetrazolium chloride, was 84.8 +/- 17.4 mm3 (mean +/- SD, n = 6). CONCLUSIONS: We demonstrated a reproducible and minimally traumatic model of brain infarction induced by the thrombotic distal middle cerebral artery occlusion in rats.

Animals↗
PubMed2005

Clinical evaluation of the traditional chinese prescription Chi-Ju-Di-Huang-Wan for dry eye.

BACKGROUND: Dry eye is a very common ocular disease characterized by eye dryness and photophobia. It often influences the patient's normal life. This study evaluated the therapeutic effects of the Chinese herb, Chi-Ju-Di-Huang-Wan, for treatment of dry eye patients. It is one of the most commonly prescribed Chinese herbs used for eyes, however, there are no scientific reports about its effect. METHODS: This study included 80 dry eye patients. The patients were randomly divided into two groups. The experimental group was treated with topical eye drops and Chi-Ju-Di-Huan-Wan. The control group was given topical eye drops and placebo. All patients received tear film examinations including Schirmer's test, Rose Bengal test, fluorescein stain test, break up time test and questionnaires. RESULTS: The results of the tear film tests were analysed by two independent t-tests between the two groups. No significant differences between the two groups according to the Schirmer's test were found. There was a significant difference in the Rose Bengal test at week 2 and tear break up time at week 4 (p = 0.04, 0.04, individually). CONCLUSION: Chi-Ju-Di-Huang-Wan is an effective stabilizer of tear film and decreases the abnormality of corneal epithelium. It provides an alternative choice for dry eye treatment.

Adult↗
PubMed1982

Identification of hepatic Z-protein in a marine elasmobranch, Platyrhinoides triseriata.

Previous studies were unable to identify Z-protein in elasmobranch liver with bromosulphophthalein as ligand. By using 8-anilinonaphthalene-1-sulphonate and Rose Bengal as ligands, however, we demonstrated in hepatic cytosol from Platyrhinoides triseriata an organic-anion-binding protein with gel-filtration characteristics identical with those of rat Z-protein. By comparison with pooled rat Z-protein, Pl. triseriata Z-protein had slightly lower affinity for 8-anilinonaphthalene-1-sulphonate and Rose Bengal, greatly decreased binding affinity for bromosulphophthalein and no binding activity for oleic acid or squalene. The Pl. triseriata Z-protein binding site was less hydrophobic than that of rat Z-protein. This observation may explain the differences in binding characteristics between the Z-proteins of these species.

Anilino Naphthalenesulfonates↗
PubMed2007

Evaluation of fertilizing potential of frozen-thawed dog spermatozoa diluted in ACP-106 using an in vitro sperm--oocyte interaction assay.

The aim of present study was to evaluate frozen canine semen with ACP-106 (Powder Coconut Water) using an in vitro sperm--oocyte interaction assay (SOIA). Ten ejaculates from five stud dogs were diluted in ACP-106 containing 20% egg yolk, submitted to cooling in a thermal box for 40 min and in a refrigerator for 30 min. After this period, a second dilution was performed using ACP-106 containing 20% egg yolk and 12% glycerol. Samples were thawed at 38 degrees C for 1 min. Post-thaw motility was evaluated by light microscopy and by using a computer aided semen analysis (CASA). Plasma membrane integrity and sperm morphology/acrosomal status were evaluated by fluorescent probes (C-FDA/PI) and Bengal Rose respectively. Moreover, frozen-thawed semen was analysed by a SOIA. Subjective post-thaw motility was 52.0 +/- 14.8% and it was significant higher than the total motility estimated by CASA (23.0 +/- 14.8%) because this system considered the egg yolk debris as immotile spermatozoa. Although normal sperm rate and acrosomal integrity evaluated by Bengal Rose stain was 89.6 +/- 3.1% and 94.3 +/- 3.1%, respectively, post-thaw percentage of intact plasma membrane was only 35.1 +/- 14.3%. Regarding SOIA, the percentage of interacted oocytes (bound, penetrated and bound and/or penetrated) was 75.3%. Using regression analysis, it was found significant relations between some CASA patterns and data for SOIA. In conclusion, the freezing-thawing procedure using ACP-106 was efficient for maintain the in vitro fertility potential of dog spermatozoa.

Acrosome↗
PubMedDate not supplied

Soluble collagen disks for the treatment of dry eye syndrome.

We evaluated the efficacy of soluble ocular inserts made from porcine scleral collagen for the treatment of dry eye syndrome. The 22 patients enrolled in this pilot study were asked to grade their pretreatment symptoms and to document the frequency of artificial tear administration. Schirmer tests, fluorescein and rose bengal staining, and impression cytology of the conjunctival epithelium were also performed at the initial evaluation. Patients were evaluated in this fashion at 3-4, 6-8, and 18-20 weeks after initiation of treatment. We observed statistically significant improvement in symptomatology and reduced necessity for artificial tear administration. There was no significant change in rose bengal staining or impression cytology results. (The absence of objective findings suggests the possibility of a placebo effect.) These findings indicate that soluble collagen inserts may be useful in treating dry eye syndrome.

Adult↗
PubMed1999

Action of agents on glucosyltransferases from Streptococcus mutans in solution and adsorbed to experimental pellicle.

Glucosyltransferase (Gtf) activity mediates sucrose-dependent adherence of mutans streptococci to the tooth surface, is essential for the cariogenicity of these micro-organisms, and contributes significantly to the exopolysaccharide component of the dental-plaque matrix. Clearly, agents that inhibit Gtfs could have therapeutic benefit. Here the effects of agents that inhibit Gtfs in solution and adsorbed to a surface were explored. Various classes of chemical reagents were tested for their ability to inhibit the enzymes responsible for insoluble-glucan synthesis (GtfB), insoluble/soluble glucan synthesis (GtfC), and soluble-glucan (GtfD) from Streptococcus mutans. Standard inhibition assays were done with Gtf enzyme in solution or with Gtf adsorbed to parotid saliva-coated hydroxylapatite (surface phase). Reagents tested included the metallic cations Li+, Zn2+, Cu2+, Fe2+ and Fe3+; the oxidizing compounds hypochlorite, Rose Bengal, perborate, and sodium-meta-periodate; and a panel of sugars and sugar analogues including sorbitol, xylitol, 1',4',6' trideoxy-trichloro-galactosucrose (TGS), and 1-deoxynojirimycin (dNJ). In solution, Gtf activity was inhibited significantly, at the highest concentrations tested: by the metal ions Zn2+, Cu2+, Fe2+ and Fe3+ (approx. 40-80% inhibition); by Rose Bengal and hypochlorite (approx. 80-90% inhibition); and by TGS and dNJ (approx. 50-80%). However, surface-adsorbed Gtfs displayed increased resistance to inhibition by the same metal cations and oxidizing compounds that inhibited them in solution. In contrast, both TGS and dNJ possessed similar inhibition profiles for both surface-bound Gtf and enzyme in solution. These data indicate that the nature of the inhibitor is important, and also whether the Gtf enzyme is in solution or adsorbed to saliva-coated hydroxylapatite.

1-Deoxynojirimycin↗
PubMed1993

Membrane potential can influence the rate of membrane photomodification.

Though cellular photomodification has been shown to change cellular resting membrane potential, an effect of membrane potential on the rate of photomodification has never been reported. Here we demonstrate that the rate of photomodification of potassium channels in frog atrial cells is voltage dependent. The rate of potassium channel photomodification using negatively charged Rose Bengal as the photosensitizer is about 2.5 times greater at the resting membrane potential of -70 mV compared to +40 mV. Similar results are obtained using the positively charged photosensitizer methylene blue. On the other hand, the rate of photomodified increase of leak current in the same cells does not significantly change in this voltage range with Rose Bengal as photosensitizer, but demonstrates a voltage dependence like that of potassium current when methylene blue is the photosensitizer. These observations cannot be explained based on voltage-dependent partitioning of the sensitizer, as similar effects on potassium current were obtained using either a positively charged or negatively charged sensitizer.

Animals↗
PubMedDate not supplied

Evaluation of retinoic acid ophthalmic emulsion in dry eye.

PURPOSE: An oil in water emulsion of 0.01% all-trans-retinoic acid (tretinoin) was prepared and clinically evaluated in dry eye patients. METHODS: The ophthalmic emulsion consisted of 10% of arachis oil and 90% of the hydrogel of Carbopol 940. To evaluate retinoic acid emulsion clinically, a placebo-controlled, open-labeled, randomized study was performed with 22 dry-eye patients. Symptoms were recorded before and after the treatments. The Schirmer I test, measurement of tear film break-up time (BUT), rose Bengal and fluorescein staining of cornea and conjunctiva, and mucus fern test were done. RESULTS: Retinoic acid did not improve the dryness, photophobia and foreign body sensation more than placebo. Schirmer test and BUT were significantly improved by retinoic acid treatment. Corneal and conjunctival epithelium maintained their characteristics during the use of retinoic acid, as indicated by rose Bengal and fluorescein staining. CONCLUSIONS: Ophthalmic emulsion of retinoic acid can be suggested as a promising approach for the treatment of dry eye.

Adult↗