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At least 703 records · Page 39Linked to original sources
PubMed1991

[Changes in the lysosome composition in cultured hepatocytes from 3-day-old rats with destabilization of the cytoplasmic microtubules].

The monolayer culture of 3-day-old rat hepatocytes was treated by antimicrotubule agent colchicine after activation with isoproterenol. Alterations in the amount of lysosomes with the size of 1.0-2.5 and 3-6 mkm per cell were studied with light microscope after staining with the vital dye neutral red and after identification of acid phosphatase by plumbum precipitation in the frozen preparations. It was shown that isoproterenol application (20 microM) within 1.5 h did not change significantly the number of lysosomes sized 1.0-2.5 microns but stimulated the increase in the amount of lysosomes 3-6 mkm in size. The addition of 2.5 mM colchicine, after the isoproterenol treatment, decreases greatly the number of lysosomes with the size of 1.0-2.5 and 3-6 mkm. A considerable part of the large lysosomes can be resistant to the colchicine action. It is concluded that the microtubules play an important role in lysosome functioning.

Animals↗
PubMed1997

In vitro cytotoxicity of guanylhydrazones (MGBG and PGBG) on cultured Chinese hamster ovary cells.

Guanylhydrazones, methylglyoxal bis(guanylhydrazone) (MGBG) and a new compound, phenylglyoxal bis(guanylhydrazone) (PGBG), interfering with polyamine biosynthesis have considerable potential for the use as antiparasitic and antitumor agents. The effect of these drugs on the cellular viability of Chinese hamster ovary cells was examined by in vitro neutral red assay. The time exposure and metabolic influence was studied. These compounds have a dose- and time-dependent cytotoxicity. The IC50 values were of 597.22 micrograms/ml and 1.77 micrograms/ml for MGBG after 3 and 24 h of incubation, respectively, and 380.50 micrograms/ml for PGBG after 24 h of incubation. The PGBG treatment during 3 h had no cytotoxic effect when the concentrations were lower than 3000 micrograms/ml. With the cytotoxicity assay used, we observed that the presence of S9 in cultured medium did not influence the cytotoxicity of these compounds.

Adenosylmethionine Decarboxylase↗
PubMed1998

Evaluation of the use of two human cell lines for okadaic acid and DTX-1 determination by cytotoxicity assays and damage characterization.

Two human cell lines have been used, HEp-2 and (de)differentiated Caco-2, derived from a larynx and a colon carcinoma, respectively, with the aim of evaluating and characterizing the cytotoxicity of okadaic acid (OA) and related toxins. Effects of OA and dinophysistoxin-1 (DTX-1) on cell viability (neutral red uptake) and on cell morphology/cytoskeleton structure have been observed in both cell lines, though at different time exposures and with different concentrations. The morphological alteration was detected earlier than the viability inhibition in HEp-2 cells with both toxins and in Caco-2 cells with DTX-1. HEp-2 cells have shown to be more sensitive than the intestinal cell line and thus possibly suitable for screening of contaminated samples, while Caco-2 cells could be used for further investigating the possible mechanisms involved in diarrhoeic shellfish poisoning (DSP) toxins.

Apoptosis↗
PubMed2003

Effect of ofloxacin and netilmicin on human corneal and conjunctival cells in vitro.

The purpose of this study was to compare the cytotoxic effects of the fluoroquinolone ofloxacin with that of the aminoglycoside netilmicin. Human corneal epithelial cells (HCE-T) and human conjunctival epithelial cells (Wong-Kilbourne derivative of Chang conjunctiva) were exposed to antibiotics (0.08-5.0 mg/mL) for 4 or 24 hours. Cell proliferation and viability were assessed with the MTT assay, neutral red uptake, and bromo deoxy uridine incorporation. In both cell lines, ofloxacin inhibited cell proliferation and viability. These effects were time and dose dependent. Concentrations of ofloxacin ranging from 0.4 to 2.4 mg/mL (0.04% to 0.24%) produced a 50% inhibition of proliferation and viability. In contrast, netilmicin induced no toxic effect. The differences between ofloxacin and netilmicin were highly statistically significant (p < 0.001). This finding is particularly relevant in deciding the optimal antibiotic to be applied in clinical situations in which the epithelium is compromised.

Administration, Topical↗
PubMed1976

Photodynamic inactivation in recurrent infections with herpes simplex virus.

Photodynamic inactivation with neutral red (a heterotricyclic dye) and light was evaluated in a placebo-controlled study of 170 episodes of recurrent infection with herpes simplex virus in 96 patients. The technique inactivated herpes simplex virus in vitro. However, no beneficial effect, either on the rate of resolution of herpetic lesions or on the interval to subsequent recurrences, was observed in treated patients. These negative results should not deter further investigation of photodynamic inactivation as a potential treatment for herpes simplex virus infections, but the clinical application of this form of therapy should be limited to properly controlled studies until its efficacy is proven.

Clinical Trials as Topic↗
PubMed2004

Synergistic effects of fumonisin B1 and ochratoxin A: are in vitro cytotoxicity data predictive of in vivo acute toxicity?

Contamination of food and feeds by mycotoxins is a major problem of human and animals health concern which is also extremely detrimental to economy. Mycotoxins producing moulds may produce a diversity of toxins such as aflatoxins, ochratoxins, trichothecenes, zearalenone, fumonisins, tremorgenic toxins and ergot alkaloids. Although toxicological, environmental and epidemiological studies have addressed the problem of these toxins one by one, more than one mycotoxin are found usually in the same contaminated commodities. That rises the incommensurable problem of multi-toxicosis in which the respective metabolites are also involved. These mycotoxins bear potential toxicity leading to acute and chronic effects in humans and animals, depending on species. The mechanisms that lead to toxic effects, such as immune toxicity, and carcinogenicity are complexe. The risk assessment for humans potentially exposed to multi-mycotoxins suffers very much from the lack of adequate food consumption data. Furthermore, for a given mycotoxin synergism and antagonism with other mycotoxins found in the same food commodities are not taken into account. The case of combination of ochratoxin A (OTA) and fumonisin B1 (FB1) has been addressed in the present paper with the purpose of predicting the in vivo toxicity using a simple in vitro test, i.e. neutral red uptake, in three different cell-lines, C6 glioma cells, Caco-2 cells and Vero cells. Using the equation of [ATLA 27 (1999) 957], in vivo toxicity (LD50) is in adequation with the in vitro data, (IC50 values) for both toxins as well as for the combination of 10 microM OTA and variable concentrations of FB1 (10-50 microM). A synergistic effect is prouved in vitro that is in line with some in vivo data from the literature. Such simple in vitro test may thus help predicting in vivo toxicity of combinations of mycotoxins naturally occurring in foodstuffs.

Animals↗
PubMed1978

In vitro and in vivo photosensitized inactivation of dermatophyte fungi by heterotricyclic dyes.

The ability of three heterotricyclic dyes to photosensitize dermatophyte fungi was studied with Trichophyton mentagrophytes and Microsporum gypseum. In vitro studies showed that methylene blue, neutral red, and proflavine were capable of killing these fungi when used in conjunction with broad-spectrum light. Proflavine, however, killed both fungi most rapidly and was used for further studies. Fungal killing by proflavine plus light was dependent on dye concentration, pH, light wavelength, and light intensity. Based on the in vitro studies, a treatment regimen was developed for in vivo use on experimentally infected animals. When treatment of guinea pigs inoculated with T. mentagrophytes was begun during fungal invasion, lesion formation at inoculated sites was either prevented or substantially reduced. When treatment was begun after lesion formation, however, light-plus-dyed treated sites showed only slightly faster curing than untreated sites.

Animals↗
PubMed1984

An improved colorimetric assay for T cell cytotoxicity in vitro.

An improved colorimetric assay for estimation of cytotoxic T (Tc) cells is described. The method involves staining thioglycollate-induced macrophage targets with the dye neutral red prior to addition of cytotoxic T cells and estimating macrophage survival at the end of the assay by measuring dye remaining in viable targets. The method using macrophage targets is more sensitive than the 51Cr release assay employing macrophages or a variety of other targets. It may be used to detect alloreactive and H-2 restricted Tc cells in both short-term (4 h) and long-term (24 h) assays and overcomes some variability encountered with a previously described colorimetric procedure. Furthermore, the method is cheap, fast, reliable and avoids the use of radioactivity.

Animals↗
PubMed2005

A novel in vitro exposure technique for toxicity testing of selected volatile organic compounds.

Exposure to vapours of volatile chemicals is a major occupational and environmental health concern. Toxicity testing of volatile organic compounds (VOCs) has always faced significant technological problems due to their high volatility and/or low solubility. The aim of this study was to develop a practical and reproducible in vitro exposure technique for toxicity testing of VOCs. Standard test atmospheres of xylene and toluene were generated in glass chambers using a static method. Human cells including: A549-lung derived cell lines, HepG2-liver derived cell lines and skin fibroblasts, were grown in porous membranes and exposed to various airborne concentrations of selected VOCs directly at the air/liquid interface for 1 h at 37 degrees C. Cytotoxicity of test chemicals was investigated using the MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) and NRU (neutral red uptake) assays following 24 h incubation. Airborne IC(50) (50% inhibitory concentration) values were determined using dose response curves for xylene (IC(50)=5350+/- 328 ppm, NRU; IC(50)=5750+/- 433 ppm, MTS in skin fibroblast) and toluene (IC(50)=0 500+/- 527 ppm, NRU; IC(50)=11,200 +/- 1,044 ppm, MTS in skin fibroblast). Our findings suggest that static direct exposure at the air/liquid interface is a practical and reproducible technique for toxicity testing of VOCs. Further, this technique can be used for inhalational and dermal toxicity studies of volatile chemicals in vitro as the exposure pattern in vivo is closely simulated by this method.

Air Pollutants↗
PubMed1995

Potential neurotoxicity of a novel aminoacridine analogue.

1. A class of compounds, 9-aminoacridines, have long been known to be reversible inhibitors of acetylcholinesterase (AChE-EC 3.1.1.7), the most familiar of which is 9-amino-1,2,3,4-tetrahydroacridine (Tacrine). 2. A novel aminoacridine was synthesised: -2-tertiary-butyl-9-amino-1,2,3,4- tetrahydroacridine (2tBuTHA). 3. In vitro comparisons of the acetylcholinesterase inhibitory potential and neurotoxicity compared to Tacrine were performed using a chemically differentiated neuroblastoma cell line (Neuro 2A). 2tBuTHA, but not Tacrine, was cytotoxic to the neural cell following 20 h exposure, despite being the least potent AChE inhibitor (IC80 AChE 12.53 microM +/- 1.14 s.e.m., Neutral Red Uptake IC50 9.53 microM +/- 0.98 s.e.m., MTT Reduction IC80 14.6 microM +/- 1.43 s.e.m.). 4. In vivo studies used a novel application of a five arm radial maze to assess neuropharmacological effects on working memory in control and Scopolamine (1 mg kg-1 i.p.) treated mice. There was an impairment of short term cognitive function with 2tBuTHA (15 mg kg-1 i.p.), but not Tacrine (10 mg kg-1 i.p.) which improved the Scopolamine deficit as expected. 5. This combined in vitro and in vivo data infers a neurotoxic property for the novel compound 2tBuTHA, a close structural analogue of Tacrine.

Aminoacridines↗
PubMed1993

Changes in heat shock protein 70 and ubiquitin mRNA levels in C1300 N2A mouse neuroblastoma cells following treatment with iron.

We have shown that following heat shock (42.5 degree C for 30 min), mouse-derived C1300 N2A neuroblastoma cells contain increased levels of mRNA coding for the inducible form of heat shock protein 70 and for ubiquitin. Incubation of C1300 cells with iron also induces an elevation in content of mRNAs coding for the same two proteins that can be blocked by alpha-tocopherol and desferrioxamine. Iron was shown to increase mitochondrial and lysosomal activities in differentiated C1300 N2A cultures, as shown by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and neutral red cytotoxicity assays. These responses were not initially associated with any loss of viability, as assessed by the lactate dehydrogenase release assay. These results suggest that there is production of cytoprotective heat shock proteins in response to iron-mediated cell damage, probably involving free radical generation, in neural cells. The apparent stress response of vulnerable neurones in human neurodegenerative diseases, particularly Parkinson's disease, may be induced by iron-mediated free radical production in degenerating neurones, making investigation of the mechanism of free radical-induced responses in neuronal cells of special interest.

Animals↗
PubMed1978

Fluorochrome staining of multilamellar liposomes.

Multilamellar liposomes can be stained with such fluorochromes as acridine orange, eosin Y, neutral red, and thiazine red. The liposomes are brought into a 1% solution of the fluorochrome; 5-10 minutes later they are centrifuged and washed by resuspending in water or phosphate buffered saline three times. The last pellet is resuspended and a drop studied with the fluorescence microscope (1000 x magnification). The fluorochrome is seen to be accumulated in the liposomal membranes. Acridine orange could also be trapped in the aqueous compartments of the liposomes but the trapped fluorochrome was gradually lost from the liposomes. Part of the fluorochrome, however, remained associated with the liposomal membranes for a long time. Additional experiments justify the conclusion that an equilibrium is maintained between fluorochromes in the aqueous and lipid phases.

Acridine Orange↗
PubMed1995

Comparison of ketoconazole- and fluconazole-induced hepatotoxicity in a primary culture system of rat hepatocytes.

Ketoconazole (KT) and fluconazole (FLU) are azole antifungal agents with a broad spectrum of activity against both superficial and systemic mycoses. KT is also an anticancer agent in the treatment of advanced prostate cancer. In many clinical and retrospective studies, KT has been reported to cause liver damage, i.e. chemical hepatitis. Histologic analysis of KT induced hepatotoxicity shows massive centrilobular necrosis in which the hepatotoxicity was not thought to be mediated through an immunoallergic mechanism. According to the medical literature, the pattern of hepatic injury appears to be primarily of the hepatocellular type. Because of the documented reports of KT and FLU hepatotoxicity, a cytotoxicity comparison of KT and FLU was implemented. The objective of this comparison was to evaluate the cytotoxicity of these azoles such that future mechanistic investigations of hepatotoxicity could be performed. The relative hepatotoxicity of KT and FLU was evaluated using primary cultures of postnatal rat hepatocytes. Cytotoxicity was evaluated by measuring the leakage of the cytosolic enzyme, lactate dehydrogenase (LDH), into the medium; by assessing mitochondrial reduction of 3-(4,5-dimethythiazol-2yl)-2,5-diphenyl tetrazolium bromide (MTT); by assessing lysosomal uptake of neutral red (NR); and by gross morphology (phase contrast microscopy). The cultures were exposed to various concentrations of KT (56-188 microM) for 0.5-4 h and to various concentrations of FLU (50 microM to 1.0 mM) for 0.5-6 h. There was a significant increase (P < 0.05) in LDH leakage and a large decrease in MTT reduction and lysosomal uptake of NR at 4 h for KT. One millimolar FLU had minimal effects on the LDH leakage and MTT reduction. These results demonstrate that KT is a more potent cytotoxicant than FLU; and its toxicity was expressed in a dose- and time-dependent manner.

Analysis of Variance↗
PubMed2001

The vacuole as central element of the lytic system and sink for lipid droplets in maturing appressoria of Magnaporthe grisea.

Histochemical and ultrastructural studies were carried out on a wild-type strain (Guy11) and a melanin-deficient mutant (buf1) of the rice-blast pathogen, Magnaporthe grisea (= Pyricularia oryzae), in order to investigate the destination of lipid storage reserves during appressorium development. Lipid droplets were abundant in conidia and were mobilised upon germination, accumulating in the appressorial hook which developed at the tip of each germ tube. Following the formation of a septum at the base of the nascent appressorium, one or a few closely appressed central vacuoles became established and were observed to enlarge in the course of appressorium maturation. On unyielding artificial surfaces such as glass or plastic, appressoria matured to completion within 36-48 h, by which time the enlarged vacuole filled most of the inside volume of the appressorium. Light and transmission electron microscopical observations revealed that the lipid droplets entered the vacuole by autophagocytosis and were degraded therein. Histochemical approaches confirmed the vacuole as the key lytic element in maturing appressoria. Endocytosis of a vital dye, Neutral Red, progressed via endosomes which migrated into the vacuole and lysed there, releasing their dye content into the vacuolar lumen. Furthermore, activity of the lysosomal marker enzyme, acid phosphomonoesterase, was strongly localised in the vacuole at all stages of appressorium maturation. It is therefore envisaged that vacuoles are involved in the degradation of lipid storage reserves which may act as sources of energy and/or osmotically active metabolites such as glycerol, which generate the very high turgor pressure known to be crucial for penetration of hard surfaces. On softer surfaces such as onion epidermis, appressoria of M. grisea were able to penetrate before degradation of lipid droplets had been completed.

Acid Phosphatase↗
PubMed1994

Calibration and time resolution of lumenal pH-transients in chromatophores of Rhodobacter capsulatus following a single turnover flash of light: proton release by the cytochrome bc1-complex is strongly electrogenic.

The flash-induced proton release into the lumen of chromatophores from Rhodobacter capsulatus was studied with Neutral red as pH-indicator. Calibration of the acidification jump after a single flash yielded a much larger figure, at least 0.8 units, than previously thought. A slow kinetic phase of proton release (85-90% of total) was sensitive to inhibitors of the cytochrome bc1-complex. Its half-rise time, about 10 ms, was the same as the rise time of the electrogenic reaction in the cytochrome bc1-complex that was recorded by electrochromism of carotenoids. The oxidoreduction of the two b-hemes was significantly faster (t1/2 approximately equal to 3 ms). Thus the major electrogenic event in the cytochome bc1-complex is proton and not electron transfer.

Bacterial Chromatophores↗
PubMed1996

Evaluation of the toxicological risk to humans of caulerpenyne using human hematopoietic progenitors, melanocytes, and keratinocytes in culture.

The extensive growth of Caulerpa taxifolia in the Mediterranean sea produces important quantities of bioactive secondary metabolites unable to enter the food chain. The cytotoxic effects of caulerpenyne, the major secondary metabolite from C. taxifolia, was studied in different in vitro models: skin cells, primary cultures of melanocytes and keratinocytes, immortalized keratinocytes (HaCaT and HESV), and bone marrow cells (hematopoietic progenitors CFU-GM). Typical dose-response curves from neutral red uptake and MTT assays were recorded in all models with IC50 ranging from 6 to 24 microM. Hematopoietic progenitors were more sensitive to caulerpenyne than melanocyte and keratinocyte cell lines, which could be due to their higher proliferative rate. The distribution of aggregates in colonies, macroclusters, and microclusters of hematopoietic progenitors was also altered in the presence of caulerpenyne. From our evaluation of the caulerpenyne concentrations required to result in cellular toxicity, the risks of cutaneous and/or food intoxication to humans may be considered minimal.

Cells, Cultured↗
PubMed2000

In vitro cytotoxicity of a new epoxy resin root canal sealer.

The cytotoxic effects of a new epoxy resin-based root canal sealer (AH-plus), together with those of two other commonly used endodontic sealers (AH26 and zinc oxide-eugenol), have been studied in vitro on a culture of human gingival fibroblasts. Cytotoxicity was assessed by direct incubation of sealers' extracts with the cultured fibroblasts at different time intervals. Morphological and cytotoxic effects of the sealers were evaluated microscopically and spectrophotometrically using the neutral red cytotoxicity assay. Our results demonstrated that the cytotoxic effects induced by zinc oxide-eugenol were detectable as early as 1 hr after mixing and remained at a high level until completion of the experiment (5 wk). AH26, however, induced early cytotoxic effects that lasted for 1 wk, followed by a substantial reduction in cytotoxicity. Cytotoxicity of the AH-plus was confined to the early period of experiment and was no longer detectable after 4 hr of mixing. Comparison between the results obtained for each sealer revealed significant differences at particular time intervals. Our findings suggest the potential advantage of this sealer over the other two sealers.

Bismuth↗
PubMed1982

Stimulation of S-9 fraction metabolism in rat liver and breast by vital dyes.

The vital dyes neutral red (NR), methylene blue (MB) and trypan blue (TB) induced microsomal enzymes that metabolize the chemical carcinogens 2-aminoanthracene (2AA), 6-aminochrysene (6AC) and diaminofluorene (DAF). The Ames test was used as a method of assessing the amount of microsomal enzyme activity that occurred in rat breast and liver when treated with NR, MB, and TB. Livers were more readily induced than mammary glands of midpregnant rats. Liver S-9 fraction from rats treated with vital dyes showed an increase in revertant colonies for 2AA, DAF, and 6AC over that from untreated rats whereas breast S-9 fraction from rats treated with vital dyes showed an increase in revertant colonies for 2AA over untreated.

Animals↗