Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Neutral Red”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 811 records · Page 45Linked to original sources
PubMed1996

Comparative effects of the metabolic inhibitors 2,4-dinitrophenol and iodoacetate on mouse neuroblastoma cells in vitro.

The toxic effects of two metabolic inhibitors, dinitrophenol and iodoacetic acid, were compared. Mouse neuroblastoma cell cultures (Neuro-2a) were exposed to different concentrations of the toxic compounds for 24, 48 and 72 h to study basal toxicity effects (cell proliferation by quantification of total protein content (PR) and relative neutral red uptake (RNRU) by lysosomes). The following biochemical indicators assessed in the in vitro test system were: cytosolic phosphofructokinase (PFK) and enolase (ENL) activities in glycolysis; mitochondrial succinate dehydrogenase (SDH) activity in the citric acid cycle; lysosomal beta-galactosidase (GAL) activity; and neuronal acetylcholinesterase (AChE) activity. The effects of the two metabolic inhibitors on the various indicators differed. Iodoacetic acid was found to be far more toxic than dinitrophenol to neuroblastoma cell proliferation at 24 h exposure. Though 2,4-dinitrophenol and iodoacetic acid both inhibited cell proliferation of the neuroblastoma cells, their effects on the other endpoints were opposite. Dinitrophenol was a general activator of the metabolism, particularly affecting lysosomal function. Iodoacetic acid did not significantly alter general metabolism, but considerably modified lysosomal function and AChE activity. The modification of lysosomal function of Neuro-2a cells by the two compounds was quite different: dinitrophenol increased RNRU and GAL activity, and iodoacetic acid decreased both parameters.

2,4-Dinitrophenol↗
PubMed1998

In vitro correlation between two colorimetric assays and the pyruvic acid consumption by fibroblasts cultured to determine the sodium laurylsulfate cytotoxicity.

The target of this research was to determine the cytotoxicity of sodium laurylsulfate on single-layer cultures of human fibroblasts, using two colorimetric methods (neutral red and MTT tests) and the evaluation of the pyruvic acid consumption by the cells. For the determination of the cytotoxicity by colorimetric tests, we have determined the absorbance at 540 nm using a spectrophotometer. Pyruvic acid, present in the culture medium, is the mitochondria's C3 energetic metabolite. So, a measure of the cell's consumption of pyruvic acid was developed. The reaction is as follows: Pyruvic acid + NADH --> Lactic acid + NAD+ and the enzyme employed is the LDH (lactate dehydrogenase). This method can be used to measure cytotoxicity, proliferation, and the cell's activation. The method is rapid, precise, and lacks any toxic byproduct. The absorbance was measured using a spectrophotometer at 340 nm. The consumption of pyruvic acid follows upon the fibroblast's growth. Sodium laurylsulfate cytotoxicity test after 24 h shows that the NR colorimetric test and the pyruvic acid consumption are correctly correlated (r = 0.91, alpha = 0.05). This dosage can be used to study the barrier properties of the corneocyte layer without destroying the artificial skin.

Cells, Cultured↗
PubMed1994

Heat shock response and cytotoxicity in C6 rat glioma cells: structure-activity relationship of different alcohols.

In C6 rat glioma cells, the n-alcohols methanol, ethanol, propanol, and butanol and the aromatic alcohol phenol all induce heat shock proteins (HSPs) of high molecular mass (68, 70, 90, and 110 kDa) when applied for 1 hr. The lowest alcohol concentrations that induce HSP synthesis cause about 20% cell death, as determined by neutral red assay. HSP induction thus occurs at alcohol concentrations close to the highest tolerable dose. The cytotoxicity and the potential of alcohols to induce the synthesis of HSPs increase with chain length and are correlated with the lipophilicity of the alcohols. A clear structure-activity relationship is observed for both parameters. A calculation of the putative membrane concentrations of these alcohols reveals that cytotoxic effects (50% cell death) occur at nearly the same membrane concentration (approximately 0.2 M). This also holds true for the lowest HSP 68-inducing alcohol concentrations, but at a lower concentration (approximately 0.12 M). The activities of major proteinases are affected by both heat shock and alcohols. The effects of alcohols also depend on the lipophilicity of the alcohols. Effective concentrations again are close to the highest tolerable dose. The stress reactions measured in terms of significant changes in HSP synthesis and proteinase activity provide information about the mechanisms by which toxic agents act on the cell.

Alcohols↗
PubMed1995

Anabolic-androgenic steroid-induced toxicity in primary neonatal rat myocardial cell cultures.

Recent literature reports of myocardial infarction in athletes who self-administer anabolic-androgenic steroid (AAS) and previous animal studies of the effects of AASs on the heart suggest that these drugs may be directly injurious to the myocardium. We have previously demonstrated that 100 microM testosterone cypionate (TC) inhibits all beating activity of primary neonatal rat myocardial cell cultures within 1 hr of exposure and causes significant LDH release by 4 hr of exposure, indicating a direct toxic effect of TC. The purpose of this investigation was to evaluate the effects of commonly abused AASs on primary neonatal rat myocardial cell cultures and to provide insight into early cellular changes that may lead to TC-induced toxicity. Significant LDH release was observed in 5-day-old primary myocardial cell cultures (obtained from 3-to-5-day-old Sprague-Dawley rats) exposed to 100 microM testosterone enanthate (TE), testosterone propionate (TP), and oxymetholone (O) for 4 and 24 hr and in cultures exposed to 100 microM testosterone (T) for 24 hr. Neutral red retention and MTT formazan production were significantly decreased in cell cultures exposed to 100 microM TE, TP, and O after only 4 hr of exposure, indicating a loss of viability and mitochondrial activity. However, there was no effect on viability of cell cultures exposed for 24 hr to 100 microM of a variety of other commonly abused AASs. Phase-contrast microscopy revealed complete disruption of the monolayer in cell cultures treated with 100 microM TE, TP, and O for 4 hr. Treatment of fura-2-loaded myocardial cell cultures with 100 microM TC produced no significant changes in calcium transients or baseline calcium levels for up to 13 min of exposure. These results indicate that O, T, TC, TE, and TP produce a direct toxic effect in heart cell cultures and that early (< 13 min) changes in calcium homeostasis are unlikely to participate in the mechanism of toxicity.

Anabolic Agents↗
PubMed2004

Evaluation of different toxicity assays applied to proliferating cells and to stratified epithelium in relation to permeability enhancement with glycocholate.

The purpose of the present study was to evaluate different toxicity assays for use on proliferating buccal TR146 cells and on stratified TR146 epithelium and to compare these results to the permeability enhancing effect of glycocholate (GC). Both the proliferating cells and the epithelium were exposed to different GC concentrations for 4 h. The MTS/PMS assay and neutral red (NR) retention were performed along with quantitation of ATP, lactate dehydrogenase (LDH) and extracellular protein. The toxicity was calculated as the IC50 value relative to the control. Increase in 3H-mannitol permeability across the epithelium concurrent with a decrease in the transepithelial electrical resistance (TEER) was also determined. The robustness of the epithelium was significantly higher than that of the proliferating cells (P <0.01). The ATP assay was the most sensitive assay with IC50 values of 6.4 and 11.5 mM for proliferating cells and epithelium, respectively. Intracellular LDH quantitation was the least sensitive method and extracellular LDH could not be used as a measure of toxicity partly due to interaction between LDH and GC. The effect on permeability and TEER could be correlated to the IC50 values obtained for the epithelium. The present study clearly demonstrates that for a correlation between toxicity and permeability enhancement, both studies should be performed on the epithelium.

Adenosine Triphosphate↗
PubMed2000

Haemocytes of the clam Tapes philippinarum (Adams & Reeve, 1850): morphofunctional characterisation.

Tapes philippinarum is a bivalve mollusc of the Pacific Ocean, successfully imported for human consumption into the northern Adriatic Sea (Europe). For better knowledge of its considerable adaptive ability in comparison with similar autochthonous species, a morpho-functional characterisation of its haemocytes was carried out with the establishment of short-term cell cultures (60 min at 25 degrees C). Various methods of cytochemical staining identified four cell types in the haemolymph: granulocytes (48.05% +/- 1.43), hyalinocytes (32.18% +/- 0.99), haemoblasts (18.97% +/- 0.63) and serous cells (0.8% +/- 0.19). The granulocytes, possessing cytoplasmic granules with differing dye affinity, included basophils, neutrophils and acidophils. Such granules stained vitally with Neutral Red, and correspond to lysosomes. Hydrolytic and oxidative enzymes were mainly detectable after stimulation in the presence of yeast cells. Both granulocytes and hyalinocytes were positive for alkaline phosphatase, non-specific esterase, peroxidase, and cytochrome C oxidase, whereas only granulocytes were positive for beta-glucuronidase, acid esterase, and arylsulphatase. Both cell types were competent phagocytes towards yeast and plasma had an opsonising effect. Moreover, the respiratory burst accompanied phagocytosis with superoxide anion production, recognisable through cytoplasmic deposits of formazan after treatment with nitro blue tetrazolium. Haemoblasts were small undifferentiated cells which, due to their morphology and positivity to the anti-CD34 antibody, show the typical features of stem cells. Serous cells, probably arising from Keber's gland and belonging to another differentiation pathway, contained non-sulphate acid mucopolysaccharides and play an important role in early defence mechanisms, taking part in the formation of clots.

5'-Nucleotidase↗
PubMed1999

Cytotoxic effects of extracts of compomers.

We have studied the cytotoxicity of 10 commercially available compomers. Extracts were taken in cell culture medium of non-cured, freshly-cured, and aged samples. Murine L-929 fibroblasts were exposed to the extracts for 24 h and the cytotoxicity was evaluated using dimethylthiazol diphenyltetrazolium (MTT) assay and neutral red uptake (NRU). Extracts were rated as severely, moderately, or slightly cytotoxic when the activity relative to controls was less than 30%, between 30% and 60%, or greater than 60%, respectively. Extracts of non-cured materials were rated severely toxic with both methods, with one exception. All but one freshly-cured material exhibited moderate to severe toxicity in both assays. Aged test specimens were rated moderately to severely toxic. Non-cured materials were generally more toxic than cured, with two exceptions. Aging and polishing the samples to remove non-polymerized surface film affected cytotoxicity to a varying degree. Curing reduced cytotoxicity in the MTT test from severe to moderate in 7 of 9 materials, but had relatively little effect in the NRU assay. Aging and polishing, however, had little effect on cytotoxicity evaluated by the MTT test, but markedly reduced cytotoxicity in NRU in 6 of 8 extracts. To conclude, extracts made from compomers used for dental fillings were found to be cytotoxic both before and after setting.

Analysis of Variance↗
PubMed2004

Differential c-myc expression profiles in normal human bronchial epithelial cells following treatment with benzo[a]pyrene, benzo[a]pyrene-4,5 epoxide, and benzo[a]pyrene-7,8-9,10 diol epoxide.

Bronchial epithelial cells are often exposed to airborne mutagens that have the potential to induce genetic changes involved in the development of lung cancer. Although lung tumors often display alterations in the expression of oncogenes and/or tumor suppressor genes, the role of specific chemicals and/or metabolites in causing these alterations is not well defined. The polycyclic aromatic hydrocarbon (PAH) benzo[a]pyrene (B[a]P), a by-product of combustion, is a prevalent airborne environmental mutagen and a constituent of cigarette smoke. The primary objective of this study was to compare the effect of B[a]P and two of its reactive metabolites, benzo[a]pyrene diol epoxide (BPDE or bay region epoxide) and benzo[a]pyrene-4,5-dihydroepoxide (BPE or K-region epoxide), on expression of the proto-oncogene c-myc in normal human bronchial epithelial (NHBE) cells using a quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) method. Changes in c-myc gene expression were compared with DNA adduct formation, growth inhibition, and cell-cycle progression as determined by (32)P-postlabelling, neutral red (NR), and flow cytometric analyses, respectively. None of the three test compounds altered the levels of 18S ribosomal RNA or beta-actin at the concentrations evaluated for c-myc expression, indicating that nonspecific changes in gene expression induced by cytotoxicity, for example, were not present at the concentrations evaluated. Cells exposed to B[a]P exhibited a dose-dependent increase in c-myc expression; conversely, a dose-dependent decrease in c-myc expression was observed following BPDE exposure. A marginal but concentration-dependent increase in c-myc mRNA levels was observed following exposure to the K-region epoxide. Our results demonstrated that, although B[a]P and its metabolites alter c-myc expression, the parent compound and its metabolites produce unequal and contrasting effects on the expression of this gene.

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide↗
PubMed1994

Apigenin and tangeretin enhance gap junctional intercellular communication in rat liver epithelial cells.

Two flavones, apigenin and tangeretin, were studied for their ability to modulate gap junctional intercellular communication (GJIC) in the rat liver epithelial cell line REL. Their cytotoxicity was first determined by cell density and neutral red uptake assays: neither apigenin nor tangeretin are cytotoxic at 10 and 25 microM, the concentrations used in our experiments. We then studied GJIC using the dye transfer assay and we observed that both apigenin and tangeretin enhance it, the maximum stimulation (x 1.7-1.8) being achieved at 25 microM for 24 h. When the dye transfer was enhanced, the amount of connexin 43 increased, which was demonstrated by Western blot and immunofluorescence analysis. For apigenin only, Northern blot analysis showed an accumulation of connexin 43 mRNA. In addition, the incubation of REL cells with the two compounds, for 1 or 24 h, prevented the inhibition of dye transfer by 12-O-tetradecanoylphorbol-13-acetate (1 or 10 ng/ml). The enhancement of GJIC by apigenin could be one of the major mechanisms responsible for apigenin's anti-tumour promoting action in vivo. As for tangeretin, its capacity to enhance GJIC completes its potential protective properties towards the post-initiation process.

Animals↗
PubMed1993

Cytotoxic effects of topical antimicrobial and antiseptic agents on human keratinocytes in vitro.

The cytotoxicity of commonly used antimicrobial and antiseptic agents to cultured human keratinocytes was investigated, combining a morphologic assay with a quantitative neutral red (NR) spectrophotometric assay. Main outcome criteria of response were the initial cytotoxicity (NR 90) and midpoint cytotoxicity (NR 50) and the highest tolerated dose (HTD), a concentration causing the first observed morphologic alterations. These values were compared with commonly administered clinical doses or calculated doses. Thirty-five agents were evaluated; five agents were also subjected to an assay for cell morphologic alterations, and HTD and percentage of cells taking up trypan blue and annihilation dose were assessed. These studies show that in clinical concentrations many, but not all, agents exert profound cytotoxic effects. The results suggest that under certain conditions cultured epithelial grafts may be exposed to clinical concentrations of neomycin, clindamycin, framycetin, erythromycin, gentamicin, and 0.1% solutions of povidone-iodine. Tetrachlorodecaoxygen anion complex (TCDO) should be further evaluated as a likely effective cleansing agent before culture grafting.

Anti-Bacterial Agents↗
PubMed1995

Interactive cytotoxicities of selected organic and inorganic substances to brown cells of Mercenaria mercenaria.

Toxicities of binary mixtures of Cu2+, Cd2+, benzo(a)pyrene [B(a)P] and N-ethylmaleimide (NEM) were screened using the in vitro neutral red (NR) assay to test the hypothesis that combined toxicity is more than or less than additive relative to the influence of each mixture constituent on toxicant uptake and brown cell lysosomal membrane stability. Significant cytotoxicity was observed at 25 mumol/L Cu2+, 500 mumol/L Cd2+ and 25 mumol/L NEM. B(a)P at 12 mumol/L exerted no toxicity under the conditions of the assay. Interactions between Cu2+ and NEM, between Cd2+ and NEM, and between Cd2+ and B(a)P significantly influenced brown cell survival. Comparison of observed joint toxicity with estimates made using a model of independent joint action indicates that interactive effects are less than additive in character. The 3-way interaction involving Cu2+, B(a)P, and NEM also affected brown cell survival to a statistically significant degree. However, the interactive cytotoxicity of this mixture is attributable mainly to the combined effect of Cu2+ and NEM. Results also indicate that new hypothesis and additional experimentation are needed to understand the interactive toxicity of mixture constituents.

Analysis of Variance↗
PubMed1994

Cytotoxicity of heavy metals in the human small intestinal epithelial cell line I-407: the role of glutathione.

Cytotoxicities of metal salts were determined in the intestinal epithelial cell line I-407 in microwell culture plates over 48 h using the widely utilized and accepted neutral red uptake procedure. Rank order cytotoxicities induced by the metal salts (in terms of LC50 values) were found to be HgCl2 (32 microM) > CdCl2 (53 microM) > CuCl2 (156 microM) > T12SO4 (377 microM) > Pb(NO3)2 (1.99 mM). Combined administration of the two most toxic metals at their LC50's showed that their toxicities were not additive or synergistic. The role of glutathione in determining toxicity induced by the metal salts in these cells was assessed by inhibition of its synthesis. Buthionine sulfoximine pretreatment at 1 mM, which was not toxic to the cells, caused sustained reduction in cellular glutathione content (to 13.8% after 48 h) and increased toxicities induced by HgCl2 (5.7-fold) and CuCl2 (1.44-fold) as shown by reductions in the LC50 values. Toxicity induced by the other metals remained unaffected. Administration of glutathione with either HgCl2 or CdCl2 did not protect the cells against their toxicity, and in the case of cadmium its toxicity was exacerbated. N-Acetylcysteine diminished toxicity induced by mercury but not cadmium.

Cadmium↗
PubMedDate not supplied

Evaluation of the toxicity of substituted benzthioanilides by using in vitro tests.

The cytotoxicity of 12 benzthioanilides substituted in the N-aromatic ring, and of two commercial preparations (imaverol and thiuram) for comparison, was studied with clone 81 cat cells, by determining the highest tolerated dose, and by using the neutral red uptake assay and the kenacid blue assay for total protein. The concentrations that induced 20%, 50% and 80% (IC20, IC50 and IC80) inhibition relative to controls were calculated from dose-response curves. For some compounds, rat LD50 values were also determined. All the benzthioanilide preparations showed in vitro toxicities lower than those of the fungicides imaverol and thiuram. It was confirmed that the cytotoxicities of the compounds depend on the type of substituent. The least toxic compound contained a CONHCH(2)CO(2)H substituent in the para position of the N-aromatic ring, and the most toxic compounds contained chloro and fluoro, or three chloro substituents in the anilide moiety. All the benzthioanilides tested showed fungistatic activity for dermatophytes; two of the compounds (compound 5 and compound 12) also inhibited the development of yeasts at concentrations lower than those which caused toxicity in vitro. The LD50 values and the cytotoxic concentrations in vitro were linearly related.

Anilides↗
PubMed1994

Pathobiological effects of acetaldehyde in cultured human epithelial cells and fibroblasts.

The ability of acetaldehyde, a respiratory carcinogen present in tobacco smoke and automotive emissions, to affect cell viability, thiol status and intracellular Ca2+ levels and to cause DNA damage and mutations has been studied using cultured human cells. Within a concentration range of 3-100 mM, a 1 h exposure to acetaldehyde decreases colony survival and inhibits uptake of the vital dye neutral red in bronchial epithelial cells. Acetaldehyde also causes both DNA interstrand cross-links and DNA protein cross-links whereas no DNA single strand breaks are detected. The cellular content of glutathione is also decreased by acetaldehyde, albeit, without concomitant changes in the glutathione redox status or in the content of protein thiols. Transient or sustained increases in cytosolic Ca2+ occur within minutes following exposure of cells to acetaldehyde. Moreover, acetaldehyde significantly decreases the activity of the DNA repair enzyme O6-methylguanine-DNA methyltransferase. Finally, a 5 h exposure to acetaldehyde causes significant levels of 6-thioguanine resistance mutations in an established mutagenesis model involving skin fibroblasts. The results indicate that mM concentrations of acetaldehyde cause a wide range of cytopathic effects associated with multistep carcinogenesis. The fact that acetaldehyde, in relation to its cytotoxicity, causes comparatively higher genotoxicity and inhibits DNA repair more readily than other major aldehydes in tobacco smoke and automotive emissions is discussed.

Acetaldehyde↗
PubMed2002

Derivatives of dehydrocrotonin, a diterpene lactone isolated from Croton cajucara: cytotoxicity in rat cultured hepatocytes and in V79 cells.

Derivatives of dehydrocrotonin (DHC; Compound I) with different anti-ulcerogenic properties but less toxicity were produced by reducing the cyclohexenone moiety of DHC with NaBH4 (Compound II), reducing the cyclohexenone and lactone moieties with LiAlH4 (Compound III) and transforming the lactone moiety into an amide (Compound IV) using dimethylamine. Derivatives of DHC were assayed in cultured hepatocytes and V79 fibroblasts. Three independent endpoints assays for cytotoxicity were used, namely, the DNA content, tetrazolium reduction (MTT) and neutral red uptake (NRU). Compound III was less toxic than the other DHC derivatives in both cell cultures. ICso values ranging from 250 to 600 microM were obtained for Compounds II and IV in the NRU and DNA content tests evaluated in 4-hour hepatocyte cultures. Although Compound II showed relatively low cytotoxicity in rat hepatocytes based on the NRU and DNA content assays, a very high toxicity (IC50=10 microM) was observed in the MTT test. Metabolites of Compound II in conditioned medium from 4-hour old hepatocyte cultures enhanced the MTT-reducing ability of V79 fibroblasts. The cytotoxicity of the derivatives was greater in recently isolated hepatocytes (only a 4-hour incubation for cell attachment prior to treating with the derivatives) than in hepatocytes previously cultured (24-hour incubation) before the treatment. Thus, aging reduced the cytotoxic effects of DHC derivatives in isolated hepatocytes, suggesting that P450-mediated biotransformation of such derivatives may lead to the formation of more toxic metabolites.

Animals↗
PubMed1987

Visually defined zones of focal ischemia in the rat brain.

A technique to guide brain tissue sampling in experimental focal ischemia is described. This technique allows visual identification for accurate metabolic analysis of regions with variable tissue perfusion. After 2.5 hours of unilateral middle cerebral artery occlusion (MCAO) in the rat, intravenous infusion of the vital dye, neutral red (NR), identified three distinct regions: (a) an intensely stained region consisting of the entire contralateral hemisphere and the medial ipsilateral hemisphere; (b) a blanched region in the lateral ipsilateral hemisphere; and (c) a zone of intermediately stained tissue interposed between the medial and lateral ipsilateral regions. Close regional correlation between NR staining and the iodoantipyrine intensity of adjacent brain slices suggests that NR distribution is a qualitative indicator of brain perfusion. Using the NR staining pattern to guide tissue dissection for metabolite analysis of high energy phosphates (adenosine triphosphate and phosphocreatine) and glucose-related metabolites showed that the blanched region contained ischemic tissue, whereas values from the contralateral cortex resembled those of control tissue. The intermediately stained region exhibited a gradient of metabolic perturbation. Tissue near intensely stained regions resembled control tissue, and tissue near blanched regions resembled ischemic tissue. These results confirm the concept of a metabolic penumbra. When present, it occupies only the most medial area of the intermediate region at 2.5 hours after MCAO.

Adenosine Triphosphate↗
PubMed2006

Fluoro-Jade B staining following zymosan microinjection into the spinal cord white matter.

1. The fluorescein derivate Fluoro-Jade B (FJB), which primarily labels dead or dying neurons, was used to study the acute focal inflammation in the spinal cord white matter. Inflammation was induced by microinjection of the yeast particulate zymosan to evaluate the biological effects of intraspinal macrophages activation without the confounding effects of physical trauma. 2. A single bolus of zymosan (Sigma, 75 nL) was stereotaxically injected at the thoracic level into the lateral white matter of rat spinal cord. A standard Fluoro-Jade B staining protocol was applied to spinal cord sections at 6, 12, 24 h and 2, 4 days postinjection. Neutral Red, NADPH-diaphorase, Iba1-IR, and DAPI staining protocols accomplished examination of the cells participating in the acute inflammatory response. 3. Zymosan caused formation of clearly delineated inflammation lesions localized in the lateral white matter of the spinal cord. Fluoro-Jade B stained cells in the area of inflammation were not observed at 12 h postinjection while mild FJB staining appeared at 24 h and intense staining was observed at 2 and 4 days postinjection. 4. This study shows that the acute response to zymosan-induced inflammation in the rat spinal cord white matter causes a gradual appearance of phagocytic microglia/macrophages and delayed FJB staining of the inflammatory cells. 5. FJB, a reliable marker of dying neurons, is a more universal agent than formerly believed. One possible explanation for the gradual appearance of FJB-stained cells in the area of inflammation is that specific time is required for sufficient levels of proteins and/or myelin debris of axonal origin to appear in the cytoplasm of phagocytic microglia/macrophages.

Animals↗
PubMed2002

Lysosomal response of earthworm (Eisenia fetida) coelomocytes to the fungicide copper oxychloride and relation to life-cycle parameters.

Juvenile specimens of the earthworm species Eisenia fetida Savigny, 1826, were exposed in the laboratory to a range of concentrations of the fungicide copper oxychloride. After an eight-week exposure period, the neutral-red retention (NRR) times were measured in the coelomocytes of all the worms that had matured. Life-cycle traits (survival, biomass change, cocoon production, cocoon mass, hatching of cocoons, number of hatchlings) were also monitored. Dose-related effects on NRR times, maturation, growth, and reproduction parameters were determined (one-way analysis of variance, p < 0.05). The lowest effect concentration for substrate copper (Cu) measured for the NRR time was 8.9 mg/kg (dry wt). Growth was severely affected at the highest exposure concentration (330 mg/kg substrate dry wt), at which no worms matured and no NRR times could be measured. Negative linear correlations (p < 0.05) were apparent between the NRR times and substrate and body Cu concentrations. Positive linear relationships were found between the NRR times and some life-cycle parameters (p < 0.05). This study showed that the NRR assay can indicate toxic stress due to Cu exposure at an early stage, and that NRR times can be linked to effects on certain life-cycle traits.

Animals↗