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At least 19 recordsLinked to original sources

Mechanism of stimulation of pinocytosis by trypan blue.

Trypan blue at 50 microgram/ml stimulates the pinocytic uptake of 125I-labelled PVP, but not of colloidal 198Au or formaldehyde-denatured 125I-labelled bovine serum albumin, by the 17.5-day rat visceral yolk sac incubated in vitro. Neither Trypan blue nor a combination of the dye with 125I-labelled PVP stimulated the rate of pinocytosis of liquid by the tissue. Trypan blue itself was shown to enter the yolk-sac cells by adsorptive pinocytosis. It is proposed that an interaction between Trypan blue and 125I-labelled PVP enables the latter substrate to enter the cells adsorptively by so-called 'piggy-back' pinocytosis.

Animals↗

Abrogation of tumor rejection by trypan blue.

Trypan blue treatment prevented tumor-specific rejection in three animal model systems. These included the spontaneous rejection of tumors of mice (UVT-2051) and guinea pig (line 1 hepatoma) as well as vaccine-induced rejection of a guinea pig tumor (line 10 hepatoma). Secondary immune reactions to line 10 cell challenges were not abolished by trypan blue treatment. Although trypan blue is a potent inhibitor of macrophage cytotoxicity in vitro, the mechanism by which it inhibited tumor-specific rejection has not been established.

Animals↗

The conformational characteristics of Congo red, Evans blue and Trypan blue.

The structures of the closely related bis-azo dyes Evans blue, Trypan blue and Congo red, which appeared to have different self-assembly properties and correspondingly different abilities to form complexes with amyloids and some other proteins, were compared in this work. Ab initio and semi-empirical methods were used to find the optimal structures and partial charge distributions of the dyes. The optimal structures were searched using different widely used programs. The structures of Congo red and evans blue were found to be planar, except for the torsion on the central diphenyl bond connecting the two halves of the dye. Both symmetrical parts of the molecules appeared very close to planarity. However, Trypan blue exhibits non planarity on the di-azo bonds, as well as on the central bond between the symmetrical parts of the dye. In a consequence, the non planarity of this molecule is higher than in the case of its isomer, Evans blue and Congo red as well. The extra rotation around the azo bonds extorted by the close proximity of the sulfonic groups may be the direct cause of its poor self-assembling and complexation properties versus Evans blue.

Coloring Agents↗

P2-purinoceptor antagonists: II. Blockade of P2-purinoceptor subtypes and ecto-nucleotidases by compounds related to Evans blue and trypan blue.

Effects of Evans blue and four derivatives as well as of trypan blue and four derivatives, mostly smaller fragments but two compounds with an additional ethylene bridge in the center of the molecule, were studied on contractions of the rat vas deferens elicited by alpha, beta-methylene ATP (alpha, beta-MeATP; mediated by P2X-purinoceptors), relaxations of the carbachol-precontracted guinea-pig taenia coli elicited by adenosine 5'-O-(2-thiodiphosphate) (ADP beta S; mediated by P2Y-purinoceptors), and the degradation of ATP by rat vas deferens tissue. All compounds shifted the concentration-response curve of alpha, beta-MeATP in the rat vas deferens to the right, and most compounds increased the maximum of the curve. Each member of the Evans blue series was similar in potency to the corresponding member of the trypan blue series. Where three concentrations were tested, the Arunlakshana-Schild regression was linear, and the slope did not differ from 1. The apparent Kd values were between 0.8 and 385 microM. In the guinea-pig taenia coli, only the members of the trypan blue group were relatively potent, shifting the concentration-response curve of ADP beta S to the right in a surmountable manner. In 2 of 3 cases where three concentrations were tested, the slope of the Arunlakshana-Schild regression was lower than 1. Apparent Kd values in the trypan blue group were between 5.2 and 324 microM. The removal of ATP from the medium by vas deferens tissue was decreased mainly by the members of the Evans blue group, with IC25% values between 13 and 158 (in 1 case > 1000) microM. The results indicate that the position of the sulphonate residues at the terminal naphthalene rings of these compounds hardly influences P2X purinoceptor affinity but greatly influences P2Y affinity and ecto-nucleotidase blockade. Among active compounds, apparent purinoceptor affinity and ecto-nucleotidase blockade increase with the size of the molecules up to Evans blue and trypan blue themselves; introduction of a central ethylene bridge does not result in a further gain in potency. NH01, the desmethyl derivative of Evans blue, seems to be interesting because it is the compound with the highest P2X- versus P2Y-selectivity presently available.

Adenosine Diphosphate↗

Effect of Evans blue and trypan blue on syncytia formation and infectivity of human immunodeficiency virus type I and type II in vitro.

Polyanionic compounds were used to inhibit infectivity of human immunodeficiency virus in vitro. Suramin, Evans blue, and Trypan blue were shown to inhibit syncytia formation normally observed when HIV-1-infected cells are cocultured with CD4+ cells. The inhibition was more pronounced with Evans blue than with any of the other polyanions studied. The inhibitory effect was significantly weaker in HIV-2 systems. However, the reverse transcriptase activities of both types of viruses were inhibited by Evans blue. Another polyanionic compound, phosphorothioate 28-mer cytidine homopolymer (SdC28) was shown to inhibit syncytium formation induced by HIV-1-and HIV-2-infected cells in an identical manner. Evans blue showed partial blocking of gp120 binding to CD4 in a solid-phase enzyme-linked immunosorbent assay (ELISA). These results suggest that the polyanionic dyes may exert their antiviral effects, at least in part, by interfering with the binding and fusion of HIV with susceptible T cells.

Antiviral Agents↗

Effects of trypan blue on thyroid secretion: localization of trypan blue within the colloid space and phagolysosomes of thyroid follicles.

Trypan blue was previously shown to directly inhibit thyroid secretion following TSH stimulation. Inhibition of both colloid droplet formation and thyroglobulin proteolysis was demonstrated. By observing the characteristic bright red fluorescence of the dye-protein complex, we have demonstrated that trypan blue rapidly enters the colloid space and combines with thyroglobulin. In addition, the dye in association with thyroglobulin has been demonstrated within phagosomes and phagolysosomes by centrifugation of the lysosomal (P15) fraction on both sucrose and Percoll density gradients. Lability or latency of the dye with the phagolysosomal contents was demonstrated and the dye was found in association with thyroglobulin by column chromatography. It is proposed that the complexing of trypan blue to thyroglobulin alters its attachment to specific follicular cell receptors, inhibits pinocytosis, and, thus, thyroid hormone secretion.

Animals↗

Alterations in spleen growth and sequestering function in animals receiving trypan blue.

Trypan-blue-induced alterations in spleen size and sequestering activity were studied in rats as possible experimental counterparts of the splenic enlargement and dysfunction associated with lymphomas in man. The spleen was examined radiographically and histologically following the i.v. administration of tantalum particles in animals receiving 4-17 s.c. injections of trypan blue at fort-nightly intervals and in control animals. Spleen size and sequestering activity increased in all rats receiving trypan blue. Histological studies demonstrated that an increase in mechanical filtration was responsible for the dysfunction. Tantalum was deposited extracellularly in perifollicular marginal zones in both experimental and control animals but these areas were much more prominent in experimental animals, contained more tantalum and retained the particles for longer periods.

Animals↗

Direct effects of trypan blue on thyroid secretion.

Trypan blue directly inhibited in vitro thyroid secretion (butanol soluble 125I release to the media) induced by both thyroid stimulating hormone (TSH) and dibutyryl cAMP. Intracellular colloid droplet counts were also decreased. Inhibition was directly proportional to dye concentration and could be overcome by supramaximal TSH and dibutyryl cAMP. Inhibition could be observed as early as 20 min of incubation, was not increased by preincubation, and could even be demonstrated after TSH in vivo. Trypan blue, in vivo, produced similar inhibition of thyroid secretion. Incubation of 125I-thyroglobulin with lysosomal enzymes revealed inhibition with much lower concentrations of dye. Inhibition of lysosomal enzyme(s) would not appear to explain the marked decreases in colloid droplets, and this may represent two separate effects of trypan blue on thyroid secretion.

Animals↗

Inactivation of measles and herpes simplex viruses by trypan blue.

The dye trypan blue inhibits infection of cells in tissue culture by measles and herpes simplex viruses at concentrations in excess of 0.01% (w/v) principally by direct inactivation of the virions. The inactivation is temperature-dependent (10(4) times greater at 37 degrees C than at 4 degrees C) and its mechanism appears to include inhibition of virus adsorption to cells.

Animals↗

Accumulation of DNA, RNA, and protein by cultured rat embryos following maternal administration of a teratogenic dose of trypan blue.

The effect of trypan blue on the rates of DNA, RNA, and protein accumulation in rat embryos was investigated using a system of whole embryo culture. Embryos were isolated on the 10th gestational day (sperm day = Day 0) either 4 or 24 hr after maternal injection. The cultivation period was for 24 or 42 hr. The rate of DNA and RNA accumulation in the embryo following either a 4- or 24-hr previous maternal treatment was significantly decreased at at both 24 and 42 hr of cultivation. Protein values were significantly reduced after a 4-hr maternal exposure, but only slightly reduced after a 24-hr maternal exposure. No significant differences in DNA, RNA, or protein content were observed at the beginning of cultivation, following either a 4- or 24-hr previous maternal treatment. The percentage of embryos establishing a visceral yolk sac circulation was greatly reduced, and this supports the conclusion that trypan blue interferes with yolk sac function. The most obvious teratogenic insults include failure of neuropore closure, retention of a dorsiflexed position, and cranial edema.

Animals↗

Establishment and optimization of a flow cytometric method for evaluation of viability of CD34+ cells after cryopreservation and comparison with trypan blue exclusion staining.

BACKGROUND: Trypan blue exclusion staining is probably the most frequently applied method (Method I) for assessment of viability in peripheral blood progenitor cell grafts after cryopreservation. Alternatively, a flow cytometry-based method (Method II) was established and optimized. STUDY DESIGN AND METHODS: In a first series of 22 autologous apheresis products, the influence of duration of antibody staining and red cell (RBC) lysis on viability was investigated. In a second series of 21 autologous and 1 allogeneic apheresis products, the effect of omitting the RBC lysis was evaluated. On the basis of the results of the first two series, 155 autologous and 57 allogeneic apheresis products were analyzed with Method I and the now optimized Method II. RESULTS: Halving the incubation times did not influence the viability of CD45+ or CD34+ cells. Omission of RBC lysis resulted in a significantly (p = 0.022) increased median viability of CD45+ cells (75.8% vs. 71.0%) without any influence on CD34+ cells. In the third series, the median viability of CD34+ cells (96.9%) was significantly (p < 0.0001) higher compared with the viability of CD45+ cells (76.2%) and the viability determined by Method I (86.5%). CONCLUSION: The viability of CD34+ cells was significantly higher compared with the viability of all white blood cells. The presented cytometry-based method is superior to the standard trypan blue method regarding the number of analyzable cells and documentation, regarding observer independence and standardization; it allows the analysis of the cells of interest for transplantation after minimal sample manipulation.

Antigens, CD34↗