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PubMed1998

Evaluation of a GPIIb/IIIa antagonist YM337 in a primate model of middle cerebral artery thrombosis.

We compared the antithrombotic effect of anti-GPIIb/IIIa antibody Fab fragment YM337 with that of a thromboxane A2 synthetase inhibitor, sodium ozagrel. With the monkeys under halothane anesthesia, the right middle cerebral artery was observed via a transorbital approach without cutting the dura mater. Photoillumination (wavelength 540 nm) was applied to the middle cerebral artery, and then rose bengal (20 mg kg(-1)) was administered intravenously. The experimental drugs were intravenously injected 15 min before rose bengal injection and followed by continuous infusion for 3 h after dye injection. The thrombotic occlusion induced by this photochemical reaction in monkey middle cerebral artery was reproducible. YM337 significantly prolonged the time to first occlusion and the total time of arterial patency during the 3-h observation period after dye injection. In contrast, sodium ozagrel had no significant effect. YM337 but not sodium ozagrel significantly inhibited ex vivo ADP-induced platelet aggregation. However, while sodium ozagrel significantly inhibited the thromboxane B2 generation accompanying arachidonic acid-induced platelet aggregation, YM337 had no effect on this variable. Neurological deficit in the YM337-treated animals was significantly milder than that in the control group. The area of infarct in the YM337 treatment animals was smaller than that in the control group. The novel selective GPIIb/IIIa antagonist YM337 was effective in ameliorating the decrease in patency of the middle cerebral artery and reducing the area of cerebral infarction in monkeys.

Animals↗
PubMed1972

The enzymic degradation of L-serine O-sulphate. Mechanism of the reaction.

1. During the enzyme-catalysed degradation of l-serine O-sulphate no exchange occurs between the hydrogen atom attached to the alpha-carbon atom of the substrate and the tritiated water of the incubation medium. 2. The participation of an intermediate carbanion has been demonstrated in the degradation by performing the reaction in the presence of tetranitromethane. 3. Photo-oxidation of the enzyme in the presence of Rose Bengal led to a rapid inactivation of enzyme with the concomitant loss of four histidine residues/molecule. 4. Rose Bengal was also a non-competitive inhibitor of the enzyme.

Ammonia-Lyases↗
PubMed1999

Oxidation of human catalase by singlet oxygen in myeloid leukemia cells.

Catalases are oxidized by singlet oxygen giving rise to more acidic conformers detected in zymograms after electrophoresis in polyacrylamide gels. This shift in catalase mobility can be indicative of singlet oxygen production in vivo. Catalase from human cells, as from many organisms, is susceptible to in vitro modification by singlet oxygen. Human myeloid leukemia (U937) cells were treated under different stress conditions and catalase activity and its electrophoretic mobility was monitored. The U937 cells were found to have high levels of catalase activity, as compared to cultured fibroblasts, and to be very resistant to oxidative stress. Hydrogen peroxide did not modify the electrophoretic mobility of catalase, even at doses that produced cell damage. Conditions that primarily generate superoxide, such as treatment with paraquat or heat shock, also failed to modify the enzyme. In contrast, photosensitization reactions using rose Bengal gave rise to a more acidic conformer of catalase. Singlet oxygen quenchers prevented catalase modification by rose Bengal and light. The growth medium had a photosensitizing activity. Catalase was not modified in cells illuminated in phosphate buffer but was modified in cells illuminated in phosphate buffer containing riboflavin. Intense light per se also generated a slight shift in the electrophoretic mobility of catalase. Ultraviolet light (350 or 366 nm) did cause a change in catalase, but to a less acidic catalase conformer, indicating other modifications of the enzyme. The main effect of photosensitization with methylene blue was crosslinking of the enzyme, although some shift to acidic conformers was observed at a low concentration of the photoactive compound. Results indicate that catalase can be modified by singlet oxygen generated intracellularly, even though the enzyme is predominantly inside peroxisomes. Under some photosensitization conditions, catalase modification can be used as a marker to detect intracellular singlet oxygen.

Animals↗
PubMed1986

Treatment of keratoconjunctivitis sicca with Lacrisert.

A double-blind cross-over comparative study on the effect of Lacrisert versus oculoguttae viscosae and saline has been conducted. Lacrisert is a soluble solid tear substitute which is applied once-a-day. The object was to compare the safety and patient acceptability of Lacrisert and oculoguttae viscosae, and to compare the efficacy of Lacrisert to oculoguttae viscosae and saline. Twenty-nine patients suffering from primary Sjögren's syndrome (Copenhagen criteria), 2 male and 27 female, age: median 56 years, range 32-79 years. Lacrisert was applied once-a-day, and saccharose crystals of similar appearance was used as placebo. All patients were treated for 4 periods of 3 weeks: Lacrisert + saline versus placebo + saline and Lacrisert + oculoguttae viscosae versus placebo + oculoguttae viscosae. The patients were examined before entering the study and after each of the 4 periods using: slit-lamp, corneal sensitivity, BUT, Schirmer-1-test and Rose-bengal staining. Comparing Lacrisert to saline a significant improvement in BUT (p less than 0.01) and in Rose-bengal staining (p less than 0.01) was found, whereas other patients were unchanged. Comparing Lacrisert with oculoguttae viscosae no significant difference in clinical parameters could be detected. The patients complained of minor discomfort. Most prominent was a tendency to loose the Lacrisert due to an inadequate solubilisation of the rod caused by the lack of aqueous tears. We conclude that Lacrisert may help patients with primary Sjögren's syndrome provided that they have some secretory capacity left in the tear glands.

Cellulose↗
PubMedDate not supplied

[An evaluation of the effectiveness of laboratory diagnostic methods for brucellosis].

The diagnostic value of bacteriological and serological methods for the laboratory diagnosis of brucellosis was studied. In the analysis of milk and cheese specimens Brucella cultures were isolated and differentiated as B.melitensis, biovar I, and B.abortus, biovar 4. In 25.6% of cases B.melitensis culture, biovar 1, was isolated from the blood of persons suspected for brucellosis. The isolation of B.melitensis culture from milk showed that this infective agent migrated from small animals to cattle, which was indicative of a high risk of human infection in the state of Zacatecas, Mexico. The comparative evaluation of serological diagnostic methods (the agglutination test in test tubes, Huddleson's slide test, the acidic rose bengal test and the 2-mercaptoethanol test) showed high sensitivity of rapid tests (Huddleson's test and the rose bengal test in 93.7% and 87.9% of cases respectively). The 2-mercaptoethanol test which gave positive results in 63.8% of cases provided additional information characterizing the course of infections process.

Animals↗
PubMed1986

Efficient singlet oxygen inactivation of firefly luciferase.

Firefly luciferase is inactivated by singlet oxygen at near diffusion controlled rates, 1.9 X 10(9) M-1 s-1, based on direct comparison with the oxidation of L-histidine. The inactivation kinetics are multiphasic. Inactivation is inhibitable by NaN3. Surface-separated-sensitizer (SSS) system in which singlet oxygen is produced above an air gap separating the reaction solution from the Rose Bengal sensitizer, ensuring only Type II reactions, was compared with a Sensitox II system in which the polymer bound Rose Bengal is contained in the reaction solution and both Type I and Type II reactions can occur. A slight stabilization is afforded by MgSO4.

Histidine↗
PubMed1997

Stimulating effect of xanthene dyes on immunoglobulin produced in vitro by rat spleen lymphocytes.

The effects of food additives on immunoglobulin produced in rat splenic lymphocytes were examined. The xanthene dye, Rose Bengal, enhanced IgE production, while inhibiting the production of IgG and IgM, at 50 microM. Among the xanthene dyes, Rose Bengal having 4 iodine and 4 chlorine atoms exerted the highest Ig production-regulating activity in splenocytes, and dihalogenated fluorescein, a diiodo compound, exerted similar activity, while the dichloro and dibromo compounds did not. These results suggest that halogen atoms, especially the iodine atom, in xanthene dyes play an important role in regulation of Ig production.

Animals↗
PubMed2005

[Autologous serum utilization in patients with lacrimal hyposecretion and persistent epithelial defects of cornea--clinical study].

Autologous serum was supposed to be efficient in the treatment of keratoconjunctivitis sicca and persistent epithelial defects. 14 of 24 eyes of patients with keratoconjunctivitis sicca improved fluorescein and rose Bengal scores. 3 of 6 eyes with persistent epithelial defects healed completely after this treatment and 2 improved fluorescein and rose bengal scores. All patients had been unsuccessfully treated with conventional treatment. Allergic or immune adverse reactions were not present and economical benefit was appreciated by some patients with poor socioeconomic condition.

Adult↗
PubMed1977

Treatment of keratoconjunctivitis sicca with liquid paraffin or polyvinyl alcohol in double-blind trial.

Twenty-two eyes affected with keratoconjunctivitis sicca have been subjected to double trials using the double-blind randomized cross-over technique. Polyvinyl alcohol in a 5% concentration improved the condition whereas liquid paraffin aggravated it, as estimated subjectively, and by the measurement of the break up time of the corneal film, and, finally, by the scoring of rose bengal vital staining of exposed cornea and conjunctiva. The conclusion is drawn that oil is contraindicated in keratoconjunctivitis sicca, and that rose bengal scoring (modified Bijsterveld method) is a valuable aid in control of the sicca syndrome.

Adult↗
PubMed1978

[Comparison of methods for the quantitative detection of moulds in foods. III. Comparison of different culture media for the mould plate count (author's transl)].

The mould plate count and taxonomical grouping of the mould flora of 9 moulded non-fluid foods were compared on totally 12 different media, which are commonly used for the detection of moulds.--In contrary to the bacteriological practice, moulds should only be counted on plates with 20-50 colonies, as on plates covered densely with a mixed mould flora, simple colonies are often no more detectable. Mould plate counts of all tested products were in the same range on all media with a sugar content of 1-2% (dextrose, maltose). With fruits and vegetables, less colonies were isolated on the Sugar Free Agar, Yeast Extract Agar and Aspergillus Differential Medium than on the other media. On those three media and on Plate Count Agar sporulation of many fungi was rather inhibited. On Aspergillus Differential Medium, on which Aspergillus flavus and Aspergillus parasiticus can be differentiated, no false positive colonies were to be seen.--On Malt Extract Agar with 7.5% NaCl the hydrophilic species were supressed, the xerophilic fungi were represented in greater number. The addition of 200 ppm rose bengal inhibited the growth of many fungi, but brought no rising of the colony count compared to Malt Extract Agar without rose bengal.--For the quantitive and qualitative mould plate count, Malt Extract Agar with 50 ppm Chloramphenicol and Chlortetracycline is recommended. For the detection of xerophilic moulds 7.5% NaCl should be added to that medium.--A proposition for the proceeding for the detection of moulds in foods is given.

Culture Media↗
PubMed1994

Effect of 21-aminosteroid lipid peroxidation inhibitor, U74006F, in the rat middle cerebral artery occlusion model.

The aim of this study was to evaluate the effect of 21-aminosteroid lipid peroxidation inhibitor, U74006F, on ischaemic brain tissue damage using the rat middle cerebral artery occlusion model. Under anaesthesia, the left middle cerebral artery was exposed without cutting the dura mater via a subtemporal craniotomy, under an operating microscope. Photo-illumination (wave length; 540 nm) was applied to the middle cerebral artery and then rose bengal (20 mg/kg) was administered intravenously. The middle cerebral artery was completely occluded by thrombus about 6 min after the administration of rose bengal. U74006F (1.0 mg/kg) was then injected intravenously just after the cessation of illumination. Twenty four hours after the operation, the extent of ischaemic damage was measured by magnetic resonance imaging technique. After measuring the extent of ischaemic damage, the brain was immediately removed from animals treated with or without U74006F for determination of lipid peroxidation, and the generation of free arachidonic acid in the brain. U74006F significantly (P < 0.01) reduced the size of ischaemic damage. Twenty-four hours after the operation, lipid peroxidation and the concentration of free arachidonic acid in the left hemisphere (infarction side) were significantly (P < 0.05) higher than in the right hemisphere. U74006F significantly (P < 0.05) decreased the content of lipid peroxidation products and free arachidonic acid. There was a significant (P < 0.05) correlation between the extent of ischaemic damage and the concentration of lipid peroxidation products in the left hemisphere 24 h after the operation. In conclusion, U74006F might reduce the extent of ischaemic damage by inhibiting lipid peroxidation in the brain, thus minimizing oxidative damage to neural tissues.

Animals↗
PubMed1995

[An animal model for cochlear microcirculatory disorders by photochemically initiated thrombosis].

Making an ideal animal model of cochlear microcirculatory disorders is an important method in studying inner ear microcirculation. After intravenous injection of Rose bengal (30 mg/kg), the lateral wall of the cochlea of guinea pigs was illuminated with green light (wavelength: 550 +/- 20 nm). The Rose bengal photoactivatedly produces oxygen radicals and oxygen singlets, which subsequently damage the endotheliocytes to cause adhesion and aggregation of platelets in the small vessels. After the photo-illumination, cochlea blood flow reduced sharply, and CAP amplitude decreased in 10 min and disappeared completely in about 20-40 min. With prolongation of time, stria vascularis, Corti's organ and spiral ganglion cells showed further disintegration due to ischemia, which resembled the histopathological changes of the cochlea with microcirculatory disorders. Therefore, this animal model may be considered an ideal one for studying the mechanisms of hearing loss that was caused by microcirculatory disorders and also for evaluating the effects of drugs on microcirculation of damaged cochlea.

Action Potentials↗
PubMed1992

Use of triple stain technique for simultaneous assessment of vitality and acrosomal status in boar spermatozoa.

The object of this study was to adapt the triple stain technique to diluted and incubated boar spermatozoa. Freshly ejaculated semen was resuspended in MR-A diluent to contain 3 x 10(7) cells/ml (diluted spermatozoa) and was subsequently capacitated (incubated spermatozoa). Experiments were conducted to show the conditions required for optimal staining quality and validation of triple stain technique. The most satisfactory staining solutions for diluted spermatozoa were 2% Trypan blue at 37 degrees C for 15 minutes, 0.8% Bismarck brown in 30% ethyl alcohol (pH 2.8) at 40 degrees C for 10 minutes and 0.8% rose Bengal in 0.1 M of Tris (pH 4.3) at 21 degrees C for 20 minutes. Satisfactory results were obtained for incubated spermatozoa with rose Bengal when the staining time was 10 minutes. Triple stain technique seemed to be a useful method for the simultaneous assessment of sperm vitality and acrosomal status; consequently, it should be valuable tool, for use in porcine in vitro fertilization systems.

Journal Article↗
PubMed2001

Controlled study of the use of autologous serum in dry eye patients.

PURPOSE: To determine the efficacy and safety of topical autologous serum as a treatment of dry eye patients. METHODS: A 2-month, prospective, single-masked, placebo-controlled study was conducted in patients with bilateral severe dry eye. One eye was randomized to receive the patient's own serum as a tear substitute, and the fellow eye received unpreserved normal saline solution as a placebo. Subjective symptoms and clinical parameters of dry eye including conjunctival impression cytology were assessed at baseline and 1 week, 1 month, and 2 months after treatment. RESULTS: Twelve dry eye patients were enrolled. Both subjective symptoms (discomfort, foreign-body sensation, dryness, and photophobia), objective signs (fluorescein and rose bengal staining and conjunctival impression cytology) improved significantly in treated eyes compared with baseline. Control eyes also had improvement in symptoms, signs, and rose bengal staining compared with baseline. Neither Schirmer test results nor tear break-up time improved in either group. The means score of all parameters were improved in both groups, and the results of conjunctival impression cytology were better in treated eyes; however, these results are not significantly different. There were no serious adverse effects observed in this study. CONCLUSIONS: There was a trend toward improvement in symptoms and signs of dry eye including cytologic changes after application of autologous serum in severe dry eye patients. However, this trend was not statistically significant. A larger scale study is warranted.

Administration, Topical↗
PubMed1995

Dyes as fungal inhibitors: effect on colony enumeration.

The effects of three dyes on the colony enumeration of nine fungal strains (including members of the Deuteromycetes and Zygomycetes) in pure and mixed cultures were investigated. Using malt extract agar as basal and control medium, the following dyes and concentrations were assayed: auramine (25 ppm), gentian violet (5 ppm) and malachite green (1 ppm). The chemicals commonly used in commercial media dichloran (2 ppm) and rose bengal (50 ppm) were included in the study as reference mould-spreading inhibitors. Higher counts were usually obtained in the media containing dichloran, rose bengal or auramine, including the control medium in the absence of chemical when the mixed-conidium inocula did not include a spreading mould. Nevertheless in most cases no significant differences were observed between them. Malachite green (1 ppm) performed mainly as a strong inhibitor of spreading moulds, only allowing adequate colony development and recoveries of both Fusarium and Aspergillus strains tested.

Antifungal Agents↗
PubMed1981

Determination of pipecurium bromide, a new non-depolarizing neuromuscular blocking agent, in human serum.

A method for the determination of 2 beta, 16 beta-bis (4'dimethyl-1'-piperazino)-3a, 17 beta-diacetoxy-5 alpha-androstane is described. After deproteinization a pipecurium bromide Rose Bengal complex is formed and extracted with chloroform from a mildly basic medium. The amount of pipecurium bromide is proportional to the amount of Rose Bengal in the complex, and the quantity of the latter can easily be determined by measuring its iodine content. The method is sufficiently sensitive to be used for the analysis of human serum.

Androstane-3,17-diol↗
PubMed2000

Vitamins C and E protect isolated cardiomyocytes against oxidative damage.

There is considerable evidence that oxygen free radicals are involved in reperfusion injury of ischemic myocardium. Epidemiologic studies showed an inverse correlation between plasma levels of alpha-tocopherol (vitamin E) and ascorbic acid (vitamin C) and mortality from ischemic heart disease. The present study examines the influence of both vitamins on the toxic effects of singlet oxygen on isolated rat cardiomyocytes. Freshly isolated cardiomyocytes from adult rats were exposed to singlet oxygen which was generated by photoactivation of the photosensitive dye rose bengal (10(-7) M). This procedure induced irreversible hypercontracture in about 95% of rod-shaped cardiomyocytes within 15 min after onset of photoactivation of rose bengal. Pretreatment with vitamin C (10(-5) to 10(-2) M) or E (10(-6) to 10(-3) M) reduced the number of hypercontracted cells after exposure to singlet oxygen in a concentration-dependent manner. Simultaneous application of both vitamins (vitamin E 10(-6) M plus vitamin C 10(-5) M or vitamin E 10(-5) M plus vitamin C 10(-4) M) revealed a marked overadditive protective effect against oxidative damage as compared with the single application of each vitamin. Our data show that alpha-tocopherol and ascorbic acid exert direct protective actions on isolated cardiomyocytes against oxidative damage and provide an overadditive effect if administered simultaneously.

Animals↗
PubMed1997

Nucleotide regulation and characteristics of potassium channel opener binding to skeletal muscle membranes.

[3H]P1075 binding to membrane preparations of rabbit skeletal muscle were observed in the presence of nucleotide triphosphates or diphosphates but not AMP, cAMP, adenosine, tripolyphosphate, or pyrophosphate. Nonhydrolyzable or poorly hydrolyzable ATP analogs inhibited MgATP-supported binding. The EC50 value for MgATP-supported binding (0.4 mM) was decreased approximately 10-fold in the presence of an ATP-regenerating system, and significant metabolism by membrane nucleotidases was confirmed by high performance liquid chromatographic analysis. [3H]P1075 bound to skeletal muscle with a Kd value of 37 +/- 3 nM and a Bmax value of 280 +/- 14 fmol/mg of protein. [3H]P1075 binding to subcellular fractions was highest in membranes enriched in T tubules. Specific binding was reversible, trypsin-sensitive, maximal at pH 8, and stereoselective for the (3S,4R)-enantiomer of cromakalim. Potassium channel openers exhibited a rank order of potency of P1075 > pinacidil > levcromakalim = BMS-180448 > nicorandil > diazoxide = BRL 38226. Fluorescein analogs (ethyleosin, phloxine B, and rose bengal) were relatively potent inhibitors of binding (Ki = 200-300 nM). The potassium channel openers cromakalim and BMS-180448 were competitive inhibitors of [3H]P1075 binding. In contrast, rose bengal and the ATP-regulated potassium channel antagonist glyburide increased the rate of [3H]P1075 dissociation in a manner consistent with noncompetitive interaction.

Adenosine Triphosphate↗