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PubMed1981

Uncoupler-reversible inhibition of mitochondrial ATPase by metal chelates of bathophenanthroline. II. Comparison with other inhibitors.

(1) Trisbathophenanthroline-Fe2+ (BPh3Fe2+)alters the hyperbolic relationship between concentration of ATP and reaction velocity of F1-ATPase to sigmoidal, with a simultaneous decrease in maximal velocity. (2) BPh3Fe2+ binds to the beta-subunit of F1 and competes with the binding of aurovertin. The reversal of this effect uncouplers in enhanced by ADP and diminished by ATP. BPh3Fe2+ also changes the hyperbolic concentration dependence of aurovertin binding to sigmoidal. (3) BPh3Fe2+ stabilizes F1 against the cold inactivation and cold dissociation in an uncoupler-reversible manner. (4) BPh3Fe2+ efficiently protects F1 against the light-induced inactivation occurring in the presence of Rose Bengal, and the effect is reversed by uncouplers. (5) The results are discussed in relation to the reaction mechanism of F1-ATPase and other enzymes catalyzing the reversible hydrolysis of pyrophosphate bonds.

Adenosine Triphosphatases↗
PubMed1975

Brucellosis: a study of five calves from reactor dams.

Five calves from Brucella abortus reactor cows bought after they had been on infected farms for two to six weeks, were reared in isolation and examined serologically for approximately six months prior to cultural and biological examination post mertem. One calf which had been serologically negative to the Rose-Bengal, serum agglutination and complement fixation tests for the last eight weeks of its life yielded a culture of Br abortus biotype 1. This serotype had also been isolated from its dam at parturition.

Agglutination Tests↗
PubMed1991

Stimulation of tear secretion and treatment of dry-eye disease with 3-isobutyl-1-methylxanthine.

We examined the effect of topically applied 3-isobutyl-1-methylxanthine (IBMX), a known secretagogue, on tear secretion and dry-eye disease in a clinical study. We found that IBMX produced a dose-dependent decrease in tear film osmolarity that was significant at 3.0 mmol/L (P less than .0005) in patients with dry-eye disease. This effect was not blocked by prior administration of proparacaine hydrochloride (P less than .05). Throughout a 4-week, open-label, vehicle-controlled study, IBMX decreased tear film osmolarity significantly, whereas vehicle alone did not. After 4 weeks, mean (+/- SEM) osmolarity in IBMX-treated eyes decreased from 325 +/- 3.2 mOsm/L to 312 +/- 1.8 mOsm/L but remained unchanged in vehicle-treated eyes (323 +/- 4.4 mOsm/L vs 320 +/- 4.2 mOsm/L). In our study, IBMX was significantly more effective than vehicle alone in decreasing rose bengal staining (P less than .02). Hence, topical IBMX stimulated tear secretion and decreased ocular surface disease in patients with dry-eye disease.

1-Methyl-3-isobutylxanthine↗
PubMed1986

Inhibitory effect of bile salts on the enterohepatic circulation of methotrexate in the unanesthetized rat: inhibition of methotrexate intestinal absorption.

The effect of conjugated and unconjugated bile salts on the intestinal absorption of methotrexate (MTX) in the unanesthetized rat was investigated using a recycling perfusion technique. We initially determined the general characteristics of MTX absorption in vivo. Absorption of low (0.5 microM) and high (6 microM) concentrations of MTX was linear with time for 60 min perfusion and occurred at rates of 0.2 and 1.65 nmol/100 cm dry length/min, respectively. Absorption of 0.5 microM MTX was pH-dependent and increased with decreasing perfusate pH. Absorption of MTX involves two processes: (1) a saturable process with a Kt of 0.98 microM, and (2) a nonsaturable diffusion process. The unconjugated deoxycholate and the conjugated taurocholate inhibited the intestinal absorption of 1 microM MTX in a concentration-dependent manner. The inhibitory effect of bile salts was reversible, and was not due to damage to the intestinal mucosa. The structural analogues folic acid and 5-methyltetrahydrofolate and the organic anions rose bengal and sulfobormophthalein were also inhibitory to MTX absorption. This study demonstrates that a variety of organic anions inhibit MTX intestinal absorption. The possible therapeutic importance of this observation is discussed.

Animals↗
PubMed1977

Photodynamic induction of a bacterial cell surface polypeptide.

The photodynamic action of several dyes on cells of a bacterium, tentatively identified as a species of Arthrobacter, resulted in remarkable stimulation of synthesis of a polypeptide 21,000 daltons in mass. This polypeptide resides on the cell surface and can be solubilized by sodium dodecyl sulfate without lysis of the cells. Chlorophyllin and rose bengal are effective in inducing synthesis of the polypeptide in proportion to their ability to sensitize the photooxidation of histidine. Etiolated cells of the alga Chlamydomonas reinhardtii y-1 excrete a substance into the medium that also sensitized the photoinduction of the polypeptide.

Arthrobacter↗
PubMed1989

Surface cell morphology of the anesthetic human cornea: a color specular microscopic study.

The technique of color specular microscopy has been utilized to study the surface cell morphology of the human cornea in 6 subjects with profound corneal anesthesia from diverse causes. In an analysis of cell area, perimeter, and shape, anesthetic corneas showed a statistically significant increase in cell size and perimeter. In addition, there was a loss of cellular outlines and overall increase in rose bengal staining of cells. We believe that these cellular microscopic changes reflect profound changes in epithelial cell turnover related to a decrease in neurohumoral transmitters known to be necessary for maintenance of normal corneal morphology and function.

Adult↗
PubMed1982

The serological response of cattle to vaccines against brucellosis, as measured by the brucellosis radioimmunoassay and other tests.

Serum sample were obtained from 281 heifers vaccinated with Brucella abortus strain 19, and from 50 heifers that had received two injections of killed B. abortus strain 45/20 adjuvant (K45/20A) vaccine. The serological response measured by the brucellosis radioimmunoassay (RIA) was compared with responses measured by other tests. The serological responses of cattle during the first weeks after strain 19 vaccination were found to give little guide to the frequency of persistent reactions. In the case of strain 19, persistent reactions were considered to be those occurring 12 or more months after vaccination. In heifers vaccinated at the recommended age, small numbers of persistent reactions were given by the RIA (four in 374 sera), the complement fixation test using warm fixation (CFTW) (six in 383) and cold fixation (one in 185), the serum agglutination test (two in 222) and the indirect haemolysis test (IHLT) (two in 369). The Rose Bengal plate test gave 74 persistent reactions in 374 sera. Five of the 50 heifers gave particularly prolonged responses to K45/20A vaccine. In these animals the RIA and IHLT remained positive for longer than the CFTW.

Agglutination Tests↗
PubMed2005

Highly efficient and photostable photosensitizer based on BODIPY chromophore.

Photosensitizers are reagents that produce reactive oxygen species upon light illumination and are commonly used to study oxidative stress or for photodynamic therapy. There are many available photosensitizers, but most have limitations, such as low photostability, structural instability, or a limited usable range of solvent conditions. Here, we describe a novel photosensitizer scaffold (2I-BDP) based on the unique characteristics of the BODIPY chromophore (i.e., high extinction coefficient, high photostability, and insensitivity to solvent environment). 2I-BDP shows stronger near-infrared singlet oxygen luminescence emission and higher photostability than the well-known photosensitizer, Rose Bengal. Unlike other photosensitizers, this scaffold is widely applicable under various conditions, including lipophilic and aqueous environments. HeLa cells loaded with 2I-BDP could be photosensitized by light illumination, demonstrating that 2I-BDP is potentially useful as a reagent for cell photosensitization, oxidative stress studies, or PDT.

Boron Compounds↗
PubMed1998

Snake-like chromatin in conjunctival cells of a population aged 30-60 years from Copenhagen City.

PURPOSE: To investigate the frequency of snake-like chromatin in the conjunctival epithelial cells (SLC) of a population from Copenhagen City. METHODS: A questionnaire was mailed to 504 persons aged 30-60 years. Persons with symptoms of ocular/oral dryness and a control group without symptoms were examined by the conjunctival imprint technique. RESULTS: According to this material the frequency of SLC in persons 30-60 years living in Copenhagen may be estimated to 47%, which is considerably higher than earlier reported. In most cases SLC was, however, occupying less than 1% of the total cell area. No significant association was found to abnormal results of the Schirmer test, tear film break up time, rose bengal score or tear film lipid layer thickness. The frequency of SLC in persons with KCS was 53%, however, only few cases of KCS were found in the material. CONCLUSION: Results indicate that SLC is a quite frequent and unspecific finding, probably reflecting chronic irritation of the ocular surface.

Adult↗
PubMed1976

Substrate specificity and modifications of the active centre of elastase-like neutral proteinases from horse blood leucocytes.

Two proteinases (2A and 2B) purified from the granular fraction of horse blood leucocytes degrade casein (Km values 12.8 and 6mg/ml respectively) with maximum activity at pH 7.4 and in the presence of 2m-urea. Urea-denatured haemoglobin, fibrinogen, albumin and resorcin/fuchsin-stained elastin are digested at a slower rate. The enzymes hydrolyse synthetic substrates of elastase, N-benzyloxycarbonyl-L-alanine 4-nitrophenyl ester (Km 0.114 and 0.178 mM) and N-acetyl-tri-L-alanine methyl ester (Km 5.55 and 0.98 mM), but they do not hydrolyse synthetic substrates of trypsin, chymotrypsin and thrombin. The examined proteinases are completely inhibited by 2 mM-di-isopropyl phosphorfluoridate and show a sensitivity to butyl and octyl isocyanates similar to that of pancreatic elastase. The pH-dependence of their photoinactivation in the presence of Rose Bengal indicates the presence of histidine in the active centre. Proteinase 2A rather insensitive to iodination by IC1 as is pancreatic elastase, whereas proteinase 2B is totally inactivated after incorporation of five iodine atoms per enzyme molecule.

Animals↗
PubMed1985

The role of bile acids in the effect of ethinylestradiol on the biliary excretion of exogenous organic anions.

Ethinylestradiol (EE) has a cholestatic effect and decreases the biliary excretion of xenobiotics. In the present experiments, pretreatment with EE did not influence the biliary excretion of bile acids. On the other hand, the effect of EE on the biliary excretion of exogenous organic anions and on bile production was not influenced by administration of taurocholic acid (TC) or by depletion of bile acids by cholestyramine (CHOL). However, phenobarbital (PB) administered simultaneously with EE prevented the depressive effects of EE on the biliary flow and biliary excretion rate of rose bengal (RB). These results indicate that bile acids do not play an important role in the effects of EE and that changes in the bile salt independent fraction might be responsible for the effects of EE pretreatment on the biliary excretion of exogenous organic anions.

Animals↗
PubMed2006

Development and validation of a competitive ELISA kit for the serological diagnosis of ovine, caprine and bovine brucellosis.

A competitive ELISA (Brucella-Ab c-ELISA) was standardized and validated for the detection of Brucella antibodies in cattle, sheep and goat sera using a monoclonal antibody (MAb 4B5A) produced against Brucella melitensis biotype 2. The specificity and sensitivity of the assay were 100% to a 67.5% cut-off point (B/Bo%). When compared with an indirect ELISA, the Brucella-Ab c-ELISA did not demonstrate cross-reactions when testing positive sera for antibodies to some Enterobacteriaceae. A comparison was made between the Brucella-Ab c-ELISA and the complement fixation and Rose Bengal tests. Results demonstrated that the Brucella-Ab c-ELISA is a valuable tool for the serological diagnosis of bovine and ovine/caprine brucellosis.

Agglutination Tests↗
PubMedDate not supplied

High light inhibits chlorophyll biosynthesis at the level of 5-aminolevulinate synthesis during de-etiolation in cucumber (Cucumis sativus) cotyledons.

Using the vascular plant Cucumis sativus (cucumber) as a model, we studied the effects of high (intense and excess) light upon chlorophyll biosynthesis during de-etiolation. When illuminated with high light (1500-1600 microE/m2/s), etiolated cucumber cotyledons failed to synthesize chlorophyll entirely. However, upon transfer to low light conditions (40-45 microE/m2/s), chlorophyll biosynthesis and subsequent accumulation resumed following an initial 2-12 h delay. Duration of high light treatment negatively correlated with chlorophyll biosynthetic activity. Specifically, we found that high light severely inhibited 5-aminolevulinic acid (ALA) synthesis. This effect partly could be because of the decrease in protein level of glutamyl-tRNA reductase (GluTR) observed. Protein level of glutamate-1-semialdehyde (GSA-AT) remained unchanged. It was also found that high light did not suppress HEMA 1 expression. Therefore, we speculated that this significant inhibition of ALA synthesis might have occurred mainly because of concomitant inactivation of GluTR and/or inhibition of complex formation between GluTR and GSA-AT. Our further observation that both methyl viologen and rose bengal similarly inhibit ALA synthesis under low light conditions suggested that reactive oxygen species (ROS) could be responsible for the inhibition of ALA synthesis in cotyledons exposed to high light conditions.

Aldehyde Oxidoreductases↗
PubMed1992

Changes in the diagnostic parameters during keratoconjunctivitis sicca therapy.

In 35 patients with mild, moderate and severe keratoconjunctivitis sicca, an association was found between treatment effect, break-up time value and Rose bengal score. Neither of these tests, used separately, was successful as a clinically valuable predictor of treatment effect as the scatter, in score points, was too large. Analysed together, however, as in a partial regression analysis, an adequate prediction is possible, which is clinically of some value.

Adult↗
PubMed1994

Outbreak of Brucella melitensis among microbiology laboratory workers.

We report on an outbreak of laboratory-acquired brucellosis involving four technicians working at a microbiology laboratory. All cases occurred in a period of 4 months. Blood cultures and the Rose Bengal test were positive for Brucella spp. in all cases. Microagglutination was positive for Brucella spp. at titers of between 1/40 and 1/160. All patients were cured after treatment.

Adult↗
PubMedDate not supplied

Quantitative assessment of human sperm acrosome reaction by using fluorescein isothiocyanate conjugated concanavalin A--comparison between highly purified acrosome reacted with non-acrosome reacted sperm.

In the present study, to achieve quantitative assessment of human sperm acrosome reaction by using fluorescein isothiocyanate conjugated Concanavalin A (FITC-Con A), the optimum conditions for microscopic observation were established by comparing the fluorescent images between the purified human sperm with the intact acrosome (non-acrosome reacted sperm; non-AR sperm) and those with exposed inner acrosomal membrane (acrosome reacted sperm; AR sperm). FITC-Con A stained not only the inner acrosomal membrane but also the other portions. These non-specific bindings were excluded by the competitive dissociation with alpha-D-mannose (Man) solution, the specific labeling of the inner acrosomal membrane with FITC-Con A was accomplished in the presence of 0.5 mg/mL Man. The influence of methanol fixation on the acrosome status were examined. The assessment of the acrosome reaction should evaluate whether the inner acrosomal membrane is exposed or masked by the plasma membrane, and methanol fixation is often employed in the histochemical stain. By methanol fixation, the fluorescent profiles of AR sperm were not altered before and after the treatment, bright fluorescence were found in the acrosomal regions. On the other hand, non-AR sperm gave weak fluorescence on the sperm heads before the treatment, the acrosomal regions fluoresced brightly after the treatment similar to those of AR sperm. Methanol fixation caused false positive results, it might denaturate the plasma membranes and facilitate permeability of FITC-Con A. FITC-Con A fluorescent stain and the modified triple stain (rose bengal stain) gave good positive correlation to evaluate AR sperm. This result concluded that the optimum conditions with FITC-Con A stain in the present procedure might be an useful tool for observation of human sperm acrosome reaction.

Acrosome↗
PubMed1978

Experimental and clinical trials of new 99mTc-labeled hepatobiliary agents.

Several new derivatives of lidocaine were synthesized and used to examine the intra- and extrahepatic bile ducts and gallbladder. Diisopropyl-IDA exhibited approximately twice the bile concentration of 131I-rose bengal during the first hour after injection. P-butyl-IDA also concentrates moderately well in the bile and has the added advantage of very low excretion into the urine (2%). It seems more effective than the other IDA derivatives at high bilirubin levels.

Bile Ducts↗
PubMed1990

Chemical probes for water oxidation cycle of photosystem II: Part 2--Effect of histidine modifying reagent on thermoluminescence peaks of spinach chloroplasts.

A modification of water oxidation complex in spinach chloroplasts by rose bengal (RB), a known histidine modifying agent, has been studied using thermoluminescence (TL) technique. The changes in the TL profiles at low concentrations of the dye are explained on the basis of alterations in the protein dynamics while those at higher concentrations of the dye are related to the oxidation of histidine residues.

Chloroplasts↗