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PubMed2000

Brilliant blue G selectively blocks ATP-gated rat P2X(7) receptors.

There are few antagonists selective for subtypes of the several P2X receptors, but these are needed to identify the receptors expressed on native cells and tissues. In particular, P2X(4) and P2X(7) receptor subunits are colocalized on immune, epithelial, and exocrine gland cells, but both are relatively insensitive to suramin and pyridoxal-5-phosphate-6-azo-2',4'-disulfonic acid derivative. In this article, we show that Coomassie Brilliant Blue G selectively inhibits P2X(7) receptors with nanomolar affinity. We measured currents in response to P2X receptor activation in HEK293 cells heterologously expressing human or rat P2X(1), P2X(2), P2X(3), P2X(2/3), P2X(4), P2X(1/5), and P2X(7) receptors. Brilliant Blue G produced a noncompetitive inhibition of rat and human P2X(7) receptors with IC(50) values of 10 and 200 nM, respectively. IC(50) values for inhibition of the other receptors ranged from 2 to >30 microM; the rat and human P2X(4) receptors showed IC(50) values of >10 and 3.2 microM. Coomassie Blue G also blocked YO-PRO1 uptake and membrane blebbing, which are uniquely associated with activation of P2X(7) receptors. Thus, Brilliant Blue G is at least 1000-fold more potent at rat P2X(7) receptors than at rat P2X(4) receptors.

Adenosine Triphosphate↗
PubMed1988

Fast and sensitive protein staining with colloidal acid violet 17 following isoelectric focusing in carrier ampholyte generated and immobilized pH gradients.

A new method is described for fast and sensitive staining of proteins following isoelectric focusing in carrier ampholyte and immobilized pH gradient polyacrylamide gels. After fixation with trichloroacetic acid the gels are stained for 5-10 min with 0.1-0.2% colloidal Serva Violet 17 (generic name: Acid Violet 17; Color Index No. 42,650) in 10% w/v phosphoric acid. After staining for only 0.5-3 min, major zones, corresponding to 100-500 ng protein, are visible without destaining on a weak background. Detection of minor components requires destaining with 3% w/v phosphoric acid for 5-80 min depending on gel thickness (120-500 microns) and type of support (fabric reinforced versus gels backed to a polyester film). For selected pH marker proteins (bovine serum albumin, carbonic anhydrase, horse myoglobin) a staining sensitivity of 1-2 ng/mm2 protein is found. Dye elution from stained fabric reinforced gels with 50% v/v dioxane-water, followed by absorbance measurements, results in a linear relationship over a range of 1-100 micrograms marker proteins. Staining with collodial Serva Violet 17 is the only method available for fast and high sensitivity and low background staining of immobilized pH gradient gels, without interference from selective dye binding in different pH ranges. Staining with the collodial dye is convenient by avoiding organic solvents with unpleasant vapors and potentially hazardous.

Colloids↗
PubMed1976

Banding of human chromosomes with basic fuchsin.

In this paper a technique is described for the banding of human metaphase chromosomes with basic fuchsin. The main characteristics of the G-banding pattern obtained with this cationic triphenylmethane dye are: the secondary constriction regions of chromosomes Nos. 1 and 16 are strongly stained, especially in the latter one; the heterochromatic area of chromosome No. 9, usually negative with most other G-banding techniques, is clearly visible as an intensely stained band adjacent to the centromere; the chromosomal outline is often very distinct, facilitating the study of the telomeres; a number of chromosomal regions with bright Q fluorescence such as the polymorphic regions of the chromosomes Nos. 3, 4, and Y also stain strongly with basic fuchsin. The basic fuchsin technique combines therefore properties of G-, C-, and Q-banding methods and seems very suitable for use in e.g., family and linkage studies. Several triphenylmethanes closely related to basic fuchsin produce similar banding patterns. The band-producing ability is, however, diminished in those dyes which contain methylated amino groups. If the methyl groups are attached to the carbon atoms at the 3-positions in the phenyl rings, band formation seems unaffected. The way in which basic fuchsin and chromatin may interact as well as the possible mechanism(s) of band formation with this dye are discussed.

Chromosomes↗
PubMed2006

UV light induced photodegradation of malachite green on TiO2 nanoparticles.

The photodegradation of malachite green (MG), a cationic triphenylmethane dye, is examined both under different pH values and amounts of TiO(2). After 15W UV-365nm irradiation for 4h, ca. 99.9% of MG was degraded with addition of 0.5gL(-1) TiO(2) to solutions containing 50mgL(-1) of the MG dye. The HPLC-PDA-ESI-MS technique was used to obtain a better understanding on the mechanistic details of this TiO(2)-assisted photodegradation of the MG dye with UV irradiation. Five intermediates of the process were separated, identified, and characterized for the first time. The results indicated that the N-de-methylation degradation of MG dye took place in a stepwise manner to yield mono-, di-, tri-, and tetra-N-de-methylated MG species generated during the processes. Under acidic conditions, the results indicated that the photodegradation mechanism is favorable to cleavage of the whole conjugated chromophore structure of the MG dye. Under basic conditions, the results showed that the photodegradation mechanism is favorable to a formation of a series of N-de-methylated intermediates of the MG dye.

Anti-Infective Agents, Local↗
PubMed1991

Permeability of dormant spores of Bacillus subtilis to malachite green and crystal violet.

The permeability of dormant spores of Bacillus subtilis to malachite green (MG) and crystal violet (CV) was examined by using potassium trichloro(eta 2-ethylene)platinum(II) (KTPt) as an electron-opaque marker for the dyes. The spores were treated with the dyes and other substances at 30 degrees C for 30 min or at 80 degrees C for 5 min. When the spores were incubated in 50 mM-MG solution at 30 degrees C and in 50 mM-CV solution at 30 degrees C or 80 degrees C, many small electron-dense precipitates, which were chemical complexes of dyes and platinum, were seen, mainly around the boundary between the inner and outer coat regions. The spores treated under the above conditions were not stained. Treatment with 50 mM-MG alone or a mixture of 25 mM-oxalic acid and 50 mM-CV at 80 degrees C made the spores stainable and dye-TPt precipitates were observed mainly in the outer pericortex region. Pretreatment with 25 mM-oxalic acid and 5% (v/v) phenol at 80 degrees C followed by 50 mM-CV treatment at 30 degrees C gave the same results as above. It was considered from these results that the inner coat itself might function as the primary permeability barrier to MG and CV, and that a secondary barrier to the dyes might exist around the cortex region.

Bacillus subtilis↗
PubMed1983

[Green urticaria].

A few minutes after a subcutaneous injection of patent blue dye, two patients developed acute urticaria with a characteristic blue green hue.

Female↗
PubMed2001

Complexities of measuring antagonist potency at P2X(7) receptor orthologs.

The ability of P2 antagonists to affect agonist-stimulated fluorescent dye accumulation in cells expressing human, rat, or mouse P2X(7) receptors was examined. Several compounds, including pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid (PPADS), which was previously thought to be a weak P2X(7) receptor antagonist, possessed high potency (nanomolar IC(50)) at human and rat P2X(7) receptors. However, there were species differences in antagonist potency with PPADS, pyridoxal 5'-phosphate (P5P), and periodate-oxidized ATP (OxATP) exhibiting 20- to 500-fold higher potency for human than for mouse P2X(7) receptors. HMA (5-(N,N-hexamethylene)amiloride) was also selective for human over rat P2X(7) receptors but potentiated responses at mouse P2X(7) receptors. Coomassie Brilliant Blue G (CBB) was a nonselective antagonist with high potency at mouse P2X(7) receptors (IC(50) approximately 100 nM). All compounds were noncompetitive antagonists, and potency could only be quantified by measuring IC(50) values. These values were similar when determined against EC(50) concentrations of ATP or 2'- and 3'-O-4(-benzoylbenzoyl)-ATP and, for most compounds, only slightly (3- to 5-fold) affected by agonist concentration. However, IC(50) values for KN62 (1-[N,O-bis(5-isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4-phenylpiperazine) and suramin, varied up to 25-fold depending upon agonist concentration. Furthermore, IC(50) values for KN62 and OxATP were 10-fold lower at 22 degrees C than at 37 degrees C, whereas IC(50) values for PPADS, P5P, suramin, and OxATP were up to 20-fold lower in NaCl than in sucrose buffer. Potency estimates for CBB and PPADS decreased 5-fold in the presence of bovine serum albumin, possibly due to protein binding. Given the species differences, and the effects of assay conditions on antagonist potency, caution must be exercised when interpreting results obtained with the available antagonists.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed2006

Biological decolorization of dye solution containing Malachite Green by microalgae Cosmarium sp.

The potential of Cosmarium species, belonging to green algae, was investigated as a viable biomaterial for biological treatment of triphenylmethane dye, Malachite Green (MG). The results obtained from the batch experiments revealed the ability of algal species in removing dye. The effects of operational parameters (temperature, pH, dye concentration and algal concentration) on decolorization were examined. Optimal initial pH was determined 9. The stability and efficiency of the algae in long-term repetitive operations were also examined. Michaelis-Menten kinetics was used to describe the apparent correlation between the decolorization rate and the dye concentration. The optimal kinetic parameters, nu(max) and K(m) are 7.63 mg dye g cell(-1)h(-1) and 164.57 ppm, respectively. All assays were conducted in triplicates.

Biomass↗
PubMed1989

An investigation of the fixation and staining of lipids by a combination of malachite green or other triphenylmethane dyes with glutaraldehyde.

Mixtures of the monocationic triphenylmethane dyes, malachite green or crystal violet, with glutaraldehyde, retained and stained phospholipid droplets in chloroplasts of leaves of Lolium multiflorum Lam. These dyes also stained trilinolenin; phosphatidic acid dioleoyl, dipalmitoyl; phosphatidylcholine dioleoyl, dilinoleoyl; and phosphatidylethanolamine dioleoyl on filter paper models. In this model system the dipalmitoyl derivatives of phosphatidylcholine and phosphatidylethanolamine did not stain well, if at all. Washing of the dyed samples with 0.5 M sodium chloride solution did not remove the colour, suggesting that the interaction is unlikely to be purely ionic. Except with trilinolenin, the colour and possibly the lipid samples were removed from the filter paper model system on washing with 100% ethyl alcohol. Other triphenylmethane dyes (methyl green, light green and fast green FCF) did not retain phospholipid droplets in tissue. Fast green did, however, stain phospholipids in the model system. Two quinone-imine dyes, neutral red and toluidine blue O, while staining phospholipids in the model system did not retain droplets on the chloroplasts but did assist in the retention and staining of cell membranes. The basis of the reaction between lipid and dye is discussed in relation to the structural formulae of the dyes and model lipids. It is possible that there is an interaction between the hydrophobic fatty acid ester side chains of the lipid and the dyes. Neither the phosphate nor the polyhydric alcohol moieties of the lipid seem to be essential for staining or retention of lipid.

Fixatives↗
PubMed2004

Biological treatment of textile dye acid violet-17 by bacterial consortium in an up-flow immobilized cell bioreactor.

AIMS: To develop a cost effective and efficient biological treatment process for small scale textile processing industries (TPI) releasing untreated effluents containing intense coloured Acid violet-17 (AV-17), a triphenyl methane (TPM) group textile dye. METHODS AND RESULTS: The samples collected from effluent disposal sites of TPI were used for selective enrichment of microbial populations capable of degrading/decolourizing AV-17. A consortium of five bacterial isolates was used to develop an up-flow immobilized cell bioreactor for treatment of feed containing AV-17. The bioreactor, operating at a flow rate of 6 ml x h(-1), resulted in 91% decolourization of 30 mg AV-17/l with 94.3 and 95.7% removal of biochemical oxygen demand and chemical oxygen demand of the feed. Comparison of the input and output of the bioreactor by UV-visible, thin layer chromatography and (1)H-nuclear magnetic resonance spectroscopy indicates conversion of the parent dye into unrelated metabolic intermediates. SIGNIFICANCE: These results will form a basis for developing 'on-site' treatment system for TPI effluents to achieve decolourization and degradation of residual dyes.

Bacteria↗
PubMed1991

Identification of dye-sensitive strains of Brucella melitensis.

Strains of Brucella melitensis biovars 1, 2, and 3 isolated in India, Italy, Kuwait, Saudi Arabia, the Federal Republic of Germany, and Zimbabwe were inhibited by thionin (20 micrograms/ml) but not by basic fuchsin (20 micrograms/ml) or safranin O (200 micrograms/ml). In other respects the strains were typical of their species and biovars. The existence of dye-sensitive strains of B. melitensis suggests that the present system of classification requires modification.

Bacterial Typing Techniques↗
PubMed2006

Patent blue: a novel vital dye in vitreoretinal surgery.

PURPOSE: To determine a novel vital dye (patent blue; Blueron) for vitreoretinal surgery in a prospective consecutive case series. METHODS: Five patients with either idiopathic epiretinal membrane (ERM; n = 2), proliferative vitreoretinopathy (n = 2), or a macular hole (n = 1) underwent a three-port pars plana vitrectomy. Patent blue assisted staining of the retinal surface followed by a consecutive peeling of the ERM (n = 4) or of the internal limiting membrane (ILM; n = 1) was performed. The main outcome measures were quality of intraoperative visualization of preretinal structures and postoperative visual acuity. RESULTS: The dye induced a moderate staining (++) of the ERM and a mild staining (+) of the ILM. Complete ERM and ILM removal was successfully achieved in all cases. A mean visual improvement of three Snellen lines was observed 6 months postoperatively. No visual field defects or visible retinal pigment epithelial changes were present 6 months postoperatively. CONCLUSION: Patent blue, a novel dye for intraocular applications, may be added as an alternative dye in chromovitrectomy.

Absorption↗
PubMed1998

A micromethod for protein assay using a light microscope.

A micromethod which utilizes a protein-dye binding reaction on an acetate membrane and a light microscope with an automatic exposure instrument has been developed for measuring small amounts of protein from limited biological materials. After discoidal cellulose acetate membranes (2.0 mm diameter), which absorbed 0.5 microliter of protein solution containing 0.2-2.0 micrograms of protein, were stained with Coomassie Brilliant Blue G-250, they were examined by a light microscope with an automatic exposure instrument. A linear relation was confirmed between the intensity of dye binding to protein and exposure time under specific optical conditions.

Animals↗
PubMed2006

[Lymphatic mapping and detection of sentinel node in early stages of cervical carcinoma].

OBJECTIVE: The aim of the study was to analyse two methods of intraoperative sentinel node detection using blue dye and blue dye with Tc99 labeled tracer in early stage of the cervical cancer. DESIGN: Prospective case observational study. SETTING: Department of Obstetrics and Gynecology, University Hospital Ostrava. METHODS: From May 2004 to September 2005, 49 patients with cervical cancer who underwent a radical surgery were included into the study. Sentinel lymph node was detected using blue dye in the first group of 23 patients and by blue dye with Tc99 in the second group of 26 patients. Intraoperative sentinel node detection was performed by visual aspection in the first group, and by visual aspection and by hand-held gamma-probe in the second group. RESULTS: Patients were divided according to stage of the disease into three subgroups FIGO IA2, FIGO IB1 and FIGO IB2. A total number of 1561 lymph node with an average 32 and 94 SLN with an average 1.9 were identified. The specific detection rate per site was 63% in the first group and 80.8% in the second group respectively. Metastatic disease was detected in 26 patients (41%) and metastatic involvement of SLN only in 11 patients (17.4%). Sensitivity and negative predictive value were 100% in both groups, false negativity was 0%. CONCLUSION: Detection of SLN by combination of blue dye and Tc99 labeled tracer has a higher detection rate of SLN than detection by blue dye alone.

Adult↗
PubMed1981

Propionic acid-metabisulphite-Schiff reagent for quick Feulgen staining.

This communication presents a method for the preparation of Schiff reagent in which N hydrochloric acid has been replaced by a low concentration of propionic acid. The result of using such a Schiff reagent indicates that the dye-reagent thus prepared is extrafast in action on acid-hydrolysed mammalian tissue sections. The intensity of nuclear colouration is also increased considerably, since the pH of the dye-reagent is 6.0. It is, therefore advocated that this newly developed Schiff reagent be used at 5 degrees C.

Animals↗
PubMed1979

Increased shelf-life and feulgen staining intensity of a modified trichloracetic acid-Schiff reagent.

Two modified formulae for the preparation of trichloracetic acid (TCA)-Schiff reagent have been presented in this communication. The modifications involve addition of 250 mg or 500 mg of trichloracetic acid per 100 ml of a 0.5% aqueous solution of basic fuchsin in place of NTCA which means 1.63 g of TCA per 10 ml of distilled water to be added to 100 ml of the dye solution. The use of TCA-Schiff reagents prepared by the modified method reveals a considerably faster Feulgen staining, a more intense colouration of the nuclei and a longer shelf-life of the dye-SO2 reagents as compared with the standard TCA-Schiff of Bloch and Godman. Possible interpretations for these phenomena have been presented.

Animals↗