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At least 793 records · Page 44Linked to original sources
PubMed1986

Mechanism of hepatotoxicity in periportal regions of the liver lobule due to allyl alcohol: studies on thiols and energy status.

The possible involvement of thiols and adenine nucleotides in the selective toxicity to periportal regions by allyl alcohol was evaluated in isolated perfused rat livers. Infusion of allyl alcohol (350 microM) for 20 min depleted hepatic glutathione content by 95% in both regions of the liver lobule yet damage was undetectable as indexed by release of lactate dehydrogenase or uptake of trypan blue. Perfusion for an additional 40 min in the absence of allyl alcohol resulted in lactate dehydrogenase release (2400 U/l) and uptake of trypan blue by 75% of hepatocytes in periportal regions of the liver lobule; however, dye was not taken up by cells in pericentral areas. Because thiol content was depleted in the undamaged pericentral area, it was concluded that thiol depletion alone cannot explain local toxicity to periportal regions by allyl alcohol. Perfusion with dithioerythritol (1.5 mM) prevented damage due to allyl alcohol totally. In contrast, addition of dithioerythritol 20 min after allyl alcohol did not prevent allyl alcohol-induced damage to periportal regions indicating that irreversible changes occur during the first 20 min which ultimately lead to damage. Fasting or pretreatment of rats with diethylmaleate (0.7 g/kg; 1 hr) to deplete glutathione decreased the T1/2 required for release of lactate dehydrogenase from 45 to 35 and 22 min, respectively. When methionine was infused into livers from diethylmaleate-treated rats, the T1/2 for release of lactate dehydrogenase by allyl alcohol was increased to 45 min. Infusion of allyl alcohol for 60 min also produced a significant decrease in ATP content and in the ATP/ADP ratio in periportal but not pericentral regions of the liver lobule.(ABSTRACT TRUNCATED AT 250 WORDS)

1-Propanol↗
PubMed1997

Protection of canine pancreatic microvascular endothelium against cold ischemic injury during preservation by the two-layer method.

BACKGROUND: Ischemic injury of the microvascular endothelium during cold preservation causes a disturbance of vascular microcirculation after reperfusion and results in graft failure. Recently we have shown that oxygenation of the canine pancreas during preservation by the two-layer method extends the period of preserved pancreatic viability. The aim of this study was to clarify the role of the oxygenation of the pancreas graft by the two-layer method in the viability of the microvascular endothelium during preservation. METHODS: After preservation of the canine pancreas by simple cold storage in Euro-Collins solution (EC) (group 1) or by the two-layer method using EC (group 2) for 48 hr, the viability of vascular endothelial cells was judged from nuclear trypan blue uptake. Pancreatic tissue perfusions were measured with a hydrogen gas clearance technique, and graft survival rates were examined after autotransplantation. In the control group, the grafts were autotransplanted without preservation (group 3). RESULTS: Graft survival rates were 0 of 5 (0%), 5 of 5 (100%), and 5 of 5 (100%) in groups 1, 2, and 3, respectively. The percentage of trypan blue-positive vascular endothelium in group 1 was significantly higher compared with group 3 (control) (26.4+/-1.7 vs. 7.4+/-4.3%, P<0.01). The two-layer method (group 2) decreased trypan blue uptake (11.3+/-3.7 vs. 26.4+/-1.7%, P<0.01). Pancreatic tissue perfusions after 2 hr of reperfusion were in inverse proportion to trypan blue uptake. Namely, pancreatic tissue perfusions in group 1 were significantly lower than group 3 (control) (45.6+/-12.8 vs. 64.5+/-20.6 ml/min/100 g, P<0.01). The two-layer method (group 2) improved pancreatic tissue perfusions (68.8+/-8.6 vs. 45.6+/-12.8 ml/min/100 g, P<0.01). CONCLUSION: We conclude that oxygenation of the pancreas during preservation by the two-layer method protects the microvascular endothelium from cold ischemic injury. Consequently, pancreatic microcirculation can be maintained after reperfusion, thus extending the period of preserved pancreatic viability.

Animals↗
PubMedDate not supplied

Which is the best test to evaluate the integrity of sperm plasma membrane?

The functional integrity of sperm plasma membrane of 87 semen samples collected from fertile and infertile men was assessed by the hypoosmotic swelling test (HOS). The results were correlated with sperm motility and supravital stains using trypan blue and eosin Y. These data indicate that sperm swelling and staining methods evaluate different properties of spermatozoa. Moreover the present report shows that staining percentages with trypan blue are significantly lower than with eosin Y.

Cell Membrane↗
PubMed1975

Phagocytic capability of the kidney: a possible mechanism for renal uptake of colloid in liver-spleen scanning.

Guinea pigs were given Trypan blue or India ink i.v. and i.m. to demonstrate whether proximal convoluted tubule cells of the normal kidney have potential phagocytic capability. Circulating colloidal particles of Trypan blue solution accumulated in these cells and the accumulation was related to the particle size, dose injection, and duration of experiment. These studies suggest an explanation for the clinical observation of renal visualization with 99mTc-sulfur colloid in patients with congestive heart failure.

Animals↗
PubMed1990

[The effect of pH, temperature and albumin content on the vitality of fetal rat liver cells during in vitro tests].

The influence of pH, temperature, and albumin content in the incubation medium on the vitality incubated hepatocytes was tested in vitro as well as the effects in the trypan-blue exclusion test. pH changes between 6.8 and 7.4 in the incubation medium have not any influence on the vitality of the hepatocytes or the trypan-blue exclusion test. Reduced vitality results occurred in an initial pH 7.8 in the incubation medium. The vital dye exclusion test is not useful in case of incubation temperature of 4 degrees C, since the results regarding the vitality of the hepatocytes are only apparently better. An addition of human serum albumin is necessary for a good survival of the hepatocytes during the incubation.

Animals↗
PubMed1982

Supravital and vital staining of diseased corneal endothelium in whole-mount preparations.

Whereas specular microscopy may provide valuable preoperative information about the status of the endothelial cells in clear corneas, in many diseased corneas that come to penetrating keratoplasty, stromal opacities or epithelial edema prevents determinations about the condition of the endothelium. A method has been developed wherein diseased-host penetrating keratoplasty specimens are maintained postoperatively in McCarey-Kaufman medium and are thereafter stained as whole mounts with alizarin red S and trypan blue to evaluate the status of the endothelial monolayer. Seventy diseased corneas were studied with this method. The alizarin red S stained the intercellular junctions, while the trypan blue stained nuclei and provided information about the viability of the endothelial cells that were studied. Unusual findings were the presence of binucleated endothelial cells in decompensated cases of iris clip pseudophakos and scattered iris stromal melanocytes on barred Descemet's membrane in cases of chemical injury, trauma, and congenital glaucoma. The method was found to be rapid, reliable, and inexpensive, particularly when compared with alternative methods of evaluation, such as scanning electron microscopy.

Adolescent↗
PubMed1978

Endoneurial mast cells in peripheral nerves of the armadillo dermis.

Electron microscopy of the dermis of the 9-banded armadillo, Dasypus novemcinctus, reveals an intimate relationship of mast cells to peripheral nerves. Mast cells are routinely found in the dermis in close proximity to nerves, and mast cells are also present within the perineurial cell sheath in the endoneurial space proper. These cells contain electron opaque granules and exhibit numerous surface folds as is characteristic of other species. The degranulation of the mast cells by injection of 0.5% aqueous trypan blue reveals sequential exocytosis of granules, similar to that described for the rat. Degranulation occurs in most dermal and endoneurial mast cells following a single intradermal injection of trypan blue. A small number of mast cells partially degranulate and a few exhibit no degranulation, suggesting a heterogeneous population. It is also noted that mitochondrial morphology differs in degranulating cells, in that they become swollen and vacuolated. The results of this study are discussed in light of previous results reported for other species, mainly at the light microscope level.

Animals↗
PubMed1984

The barrier function of the posterior capsule.

The permeability of the rabbit lens and human cataractous lens posterior capsule to epinephrine and trypan blue and the absorption of ultraviolet and visible light through the posterior capsule were studied in vitro. The posterior capsule served as a barrier to large nonelectrolytes or negative electrolytes other than trypan blue, but it did not impede epinephrine, ultraviolet or visible light.

Alprostadil↗
PubMed2001

[Capsulorhexis staining by trypan bleu in mature cataract surgery].

BACKGROUND: Capsulorrhexis may be hazardous in the absence of the red fundus reflex. Anterior capsule trypan blue staining enabled to perform the capsulorrhexis and safe phacoemulsification. MATERIAL AND METHODS: Twenty patients with mature cataract were enrolled in the study and opereted on after injection of trypan blue in the anterior chamber. RESULTS: Circular capsulorrhexis was performed in all cases followed by phacoemulsification and in-the-bag IOL implantation. In two cases hard nuclei required a manual extracapsular technique. No operative complications, and no residual of coloration was noted the day after the operation. With a follow-up of 2 to 12 months, no complications appeared (cornea, capsulorrhexis, IOP). CONCLUSION: In mature cataracts, with no red fundus reflex, capsulorrhexis can be done by anterior capsule staining.

Aged↗
PubMed1999

Infectivity of feline enteroepithelial stages of Toxoplasma gondii isolated by Percoll-density gradient centrifugation.

The infectivity of the feline enteroepithelial stages of Toxoplasma gondii isolated by Percoll-density gradient centrifugation was examined by the trypan blue dye exclusion method by assaying their penetration into feline fibroblast cells in vitro and by inoculation of the intestinal mucosa of cats. A large population of the parasites showed trypan blue dye exclusion activity. When feline fibroblast cells were inoculated with feline enteroepithelial stage parasites, no intracellular parasites were found 18 h post-inoculation. Kittens inoculated intraduodenally with 2 x 10(6) feline enteroepithelial stage parasites shed oocysts between 2 and 8 days post-inoculation. These results indicate that the isolated feline enteroepithelial stage parasites display infectivity towards enterocytes of cats and are capable of gametogenesis.

Animals↗
PubMed1979

Superoxide dismutase, catalase and scavengers of hydroxyl radical protect against the toxic action of alloxan on pancreatic islet cells in vitro.

Experiments with isolated pancreatic islets or dispersed islet cells from non-inbred ob/ob mice were performed to test the hypothesis that free radicals, notably OH., mediate the diabetogenic toxicity of alloxan. Accumulation of 86Rb+ by whole islets and exclusion of Trypan Blue by dispersed cells were used as previously validated criteria of islet-cell viability. Alloxan alone drastically inhibited the Rb+ accumulation and significantly decreased the frequency of cells excluding Trypan Blue. Enzymic scavengers of O2.- and H2O2 or non-enzymic scavengers of OH. or singlet oxygen were added to the incubation medium and tested for their ability to protect against these effects of alloxan. Superoxide dismutase, catalase, dimethyl sulphoxide, benzoate, and mannitol counteracted the effects of alloxan in both cytotoxicity assays. Significant protection of the Rb+-accumulating capacity was also afforded by butanol, caffeine, theophylline, NADH, NADPH and, to a small extent, NAD+. Urea has a poor affinity for OH. and did not protect against alloxan. No effect was obtained with the singlet-oxygen scavenger, histidine. Except for the protection by NADH and NADPH, which may be due to a direct reaction with alloxan in the medium, the results strongly support the hypothesis. beta-Cells may be particularly vulnerable to alloxan because their metabolic specialization facilitates reduction of the drug and perhaps of other substrates for O2.--yielding redox cycles.

Alloxan↗
PubMed1998

Pulmonary cell death in warm ischemic rabbit lung is related to the alveolar oxygen reserve.

BACKGROUND: If lungs could be retrieved for transplantation from non-heart-beating cadavers, the shortage of donors might be significantly alleviated. METHODS: We studied the effect of different postmortem lung conditions on pulmonary cell death. Lungs from 208 New Zealand white rabbits were flushed with trypan blue vital dye solution at intervals after circulatory arrest, fixed, and mounted for histologic examination. Pulmonary cells were judged to be viable on the basis of their ability to exclude trypan blue dye. In the control group, lungs were excised immediately after death and immersed in cold (4 degrees C) saline solution. In the other groups, cadavers were left at room temperature with lungs deflated, ventilated with room air or 100% oxygen or 100% nitrogen, or inflated with room air or 100% oxygen. RESULTS: There was a gradual increase in percentage (mean +/- SEM) of nonviable cells in the control group from 2.5%+/-0.9% (preischemic value) to 18.1%+/-2.8% at 24 hours after death (p < 0.001). In cadavers with lungs deflated, 79.7%+/-2.1% of cells were nonviable at 24 hours after circulatory arrest (p < 0.001 versus control group). In contrast, room air-ventilated cadavers showed only 21.4%+/-2.7% nonviable cells at this interval (p < 0.001 versus deflated group; not significant versus control group). Values in oxygen-ventilated animals were similar. Nitrogen-ventilated cadavers, however, had significantly more nonviable lung cells (73.8%+/-3.2%; p < 0.001 vs room air and oxygen-ventilated group, not significant vs deflated group). Oxygen-inflated lungs showed a parallel decrease in cell viability up to 4 hours after death when compared with room air-inflated cadaveric lungs, but thereafter more cells became nonviable in the latter group (11.1%+/-0.7% vs 19.6%+/-3.2% at 6 hours and 48.7%+/-7.2% vs 75.5%+/-4.6% at 24 hours, respectively; p < 0.01). CONCLUSIONS: Postmortem room air ventilation is as good as oxygen ventilation in delaying pulmonary cell death, and its effect is comparable to cold storage; nitrogen ventilation, however, is ineffective and not different from deflation; oxygen inflation will preserve ischemic cells for longer intervals as opposed to room air inflation. Therefore the alveolar oxygen reserve seems to be the critical factor to protect-the lung parenchyma from warm ischemic damage.

Animals↗
PubMed2005

Comparison of dyes for cataract surgery. Part 1: cytotoxicity to corneal endothelial cells in a rabbit model.

PURPOSE: To investigate the corneal endothelial cytotoxicity of dyes for capsule staining in cataract surgery. SETTING: Department of Ophthalmology and Institute of Clinical Medicine, College of Medicine, National Cheng Kung University, Tainan, Taiwan. METHODS: Cultured corneal endothelial cells of New Zealand white rabbits were exposed for 1 minute to 1 of the following dyes (various concentrations): indocyanine green (ICG), methylene blue (MB), gentian violet (GV), trypan blue (TB), and fluorescein sodium (FS). The degree of cell damage was determined by in vitro staining with TB and comparison with results in a control group. The effect of longer exposure (up to 10 minutes) to ICG 0.25% was also investigated. Structural changes in corneal endothelial cells after dye exposure were evaluated by light microscopy and transmission electron microscopy (TEM). RESULTS: Indocyanine green 0.25%, MB 0.20%, GV 0.01%, TB 0.40%, and FS 10% did not induce significant damage to corneal endothelial cells. Significant cytotoxicity was observed with the following or higher dye concentrations: ICG 0.50%, MB 0.50%, and GV 0.10%. Exposure to ICG 0.25% for 1 to 10 minutes showed a trend toward cytotoxicity after 10 minutes. On TEM, corneal endothelial cells that had been exposed to ICG 0.50% showed remarkable organelle swelling and disruption, electron-dense granules, and cell lysis. CONCLUSION: One minute of exposure to ICG 0.25%, MB 0.20%, GV 0.01%, TB 0.40%, and FS 10% appeared to be safe as determined by no cytotoxic effects on rabbit corneal endothelial cells in culture.

Animals↗
PubMed1994

Effect of extracellular Ca2+ on indomethacin-induced injury to rabbit dispersed gastric mucosal cells.

The effects of extracellular Ca2+ on indomethacin-induced disruption to rabbit dispersed gastric mucosal cells have been examined. Fundic mucosal cells were isolated, and cellular viability and disruption were assessed by the release of the lysosomal enzyme acid phosphatase and by trypan blue dye exclusion. Addition of the Ca2+ ionophore A23187 (12.5 and 25 microM) and indomethacin (0.1-10 microM) to the incubation medium induced concentration-dependent increases in enzyme marker release and trypan blue dye uptake in the cells. The resultant cellular disruption in response to incubation of cells with combinations of indomethacin and A23187 was not significantly different from that observed when the agents were administered separately. The degree of cell injury was reduced by removal of Ca2+ from the incubation medium. Similarly preincubation with the Ca2+ channel antagonist verapamil (1 microM) reduced A23187- and indomethacin-induced cell injury. Both A23187 (25 microM) and indomethacin (10 microM) treatments increased cytosolic Ca2+ concentration ([Ca2+]i). Pretreatment of cells with 16,16-dimethylprostaglandin E2 (0.1-10 microM) reduced both indomethacin (10 microM) induced cellular damage and the elevation in [Ca2+]i. These data suggest that indomethacin-induced disruption of gastric mucosal cells is dependent to some extent upon extracellular Ca2+. An inappropriate Ca2+ flux may contribute to indomethacin-induced damage to gastric mucosal cells.

16,16-Dimethylprostaglandin E2↗
PubMed1984

Comparison of two supravital stains in examination of human semen and in tests for cytotoxic antibodies to human spermatozoa.

Supravital staining of human spermatozoa using trypan blue was compared with eosin Y staining. The dyes were applied for determination of the percentage of "dead" spermatozoa in semen samples and for determination of complement-dependent antisperm antibody activity in serum. Supravital staining percentages with trypan blue were significantly lower than with eosin Y.

Antibodies↗
PubMed1995

In vitro evaluation of neutrophil viability after exposure to a hypotonic medium.

Separation techniques for radiolabelled leukocytes have inherent problems with contaminants (e.g. platelets and erythrocytes). Hypotonic lysis methods can eliminate the erythrocytes, but the question of neutrophil viability after an exposure to a hypotonic solution (i.e. sterile water) remains. Ficoll/ hypaque two-density gradient separation was performed on donor whole blood to obtain a pure neutrophil suspension. A timed sequence of water exposure was done for 5-100 s on the neutrophil preparations. The viability of these preparations was evaluated using flow cytometry and chemotaxis. The trypan blue staining method was used to document cell death. With water exposures ranging up to 100 s, 2.04 +/- 1.80% neutrophils exhibited cellular degradation by flow cytometry, and all samples demonstrated viable neutrophils by chemotaxis and trypan blue staining. The hypotonic medium exposure times for leukocyte separations should be less than 30 s for neutrophils to retain their viability by these in vitro techniques.

Cell Death↗
PubMed1980

Quantitative studies of cytological damage in mouse testis produced by exposure to heat.

Damage in gonadal cells produced by exposure of the hindquarters of conscious male mice to water at temperatures up to 43 degrees C was studied by measuring the percentage of testicular cells which do not exclude trypan blue 4 h after exposure, sperm count up to ten wk later, and the percentage of morphologically abnormal sperm up to ten wk later. It was found that the percentage of cells stained with trypan blue was significantly increased by exposure for 0.5 h at 41 degrees C or 2 h at 37 degrees C. Sperm count and the percentage of abnormal sperm were affected by 0.5 h at 43 degrees C or 2 h at 40 degrees, but not by lower temperatures. Since there is a high incidence of abnormal sperm after some heat treatments, the use of this assay for mutagen screening should be approached with caution.

Animals↗
PubMedDate not supplied

Transepithelial electrical resistance is not a reliable measurement of the Caco-2 monolayer integrity in Transwell.

The significance of monitoring transepithelial electrical resistance (TEER) value during the study on drug absorption through Caco-2 monolayers in Transwells was re-evaluated. TEER value was monitored before, during, and after the absorption of Streptokinase (45 KD). Four enhancers--disodium ethylenediaminetetracetate (disodium EDTA), sodium cholate (NaC), sodium taurocholate (NaTC), and sodium caprate along with alpha-hemolysin (a cell membrane pore-forming toxin)--were used to signify the outcome of this study. Modified trypan blue exclusion technique was used to examine the Caco-2 cell viability throughout the absorption studies. The enhancers at the used concentration exhibited toxic effect on the Caco-2 cells as evident from the trypan blue exclusion studies. This toxic effect was not reflected by the TEER profile because TEER value dropped after the addition of the absorption enhancers. But it came back to its initial value after the cell culture media was replaced by enhancer-free media. This toxic effect was confirmed by the antiproliferation studies on the four enhancers and alpha-hemolysin against Caco-2 cells. Therefore, we concluded that the measurement of TEER is not a reliable method to determine the absorption enhancers toxicity or integrity of the Caco-2 monolayers in the Transwells.

Adjuvants, Pharmaceutic↗