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At least 19 recordsLinked to original sources

PubMed1976

Feulgen-Naphthol Yellow S cytophotometry of liver cells. The effect of formaldehyde induced shrinkage on nuclear Naphthol Yellow S binding.

1. In isolated liver cells, fixed in 4 per cent formaldehyde (NFS) for Feulgen-Naphthol Yellow S (F-NYS) staining of DNA and protein, nuclear shrinkage increases the nuclear concentration of solids to 46 per cent (w/v) before the start of the NYS staining. 2. When a fixative mixture of methanol:acetic acid:formalin (85:5:10 by volume; MAF) is used, the concentration of nuclear solids during NYS staining remain at a physiological level of 19 per cent. 3. By exposing liver cells to NFS for 10 to 120 seconds before fixation in MAF, increasing nuclear shrinkage can be induced with increasing pretreatment in NFS. Nuclear NYS binding decreases in parallel with the decreasing nuclear volume in cells thus treated. As the shrinkage induced reduction in NYS binding may vary with the net charge of nuclear non-histone proteins, MAF fixation must be preferred for quantitative determinations of nuclear non-histone protein in F-NYS stained, isolated cells. 4. Fixation in MAF offers the same advantages as NFS fixation as regards the small loss of proteins during the Feulgen staining procedure and the excellent reproducibility of the F-NYS staining. Storage of MAF fixed cells in the fixative for a few days does not alter their F-NYS staining properties. 5. In MAF fixed, F-NYS stained cells there is no NYS binding to histone basic amino acid residues.

Acetates↗

PubMed1979

Naphthol AS-BI (7-bromo-3-hydroxy-2-naphtho-o-anisidine) phosphatase and naphthol AS-BI beta-D-glucuronidase in Chinese hamster ovary cells: biochemical and flow cytometric studies.

Conditions for the biochemical and flow cytometric assay of 7-bromo-3-hydroxy-2-naphtho-o-anisidine phosphatase and beta-D-glucuronidase activities in Chinese hamster ovary cells were studied. In the biochemical assay, the pH optimum for the phosphatase activity was pH 4.6 with a Km of 10(-5) M; the pH optimum for beta-D-glucuronidase activity was pH 5.0 with a Km of 2 x 10(-5) M. For intact cells the derived constants were 3 to 10 times higher. The rate of hydrolysis of both substrates was also examined by flow cytometry. Cellular fluorescence increased linearly for only about 15 min. Diffusion of the fluorescent product probably caused nonlinearity of the fluorescence increase and was demonstrated by mixing cells incubated with substrate with those that had not been incubated. After 15 min, cells that had not been exposed previously to product or substrate contained the fluorescent product. Cells fractionated into size classes by centrifugal elutriation also were analyzed by flow cytometry for beta-D-glucuronidase activity. The activity increased linearly with the increase in cell size corresponding to the progression from G1 through S and into G2-M phases of the cell cycle.

Animals↗

PubMed1978

The influence of dose on the pattern of conjugation of phenol and 1-naphthol in non-human primates.

1. The pattern of conjugation of phenol and 1-naphthol was investigated in several primates; three Old World species (rhesus, cynomolgus, patas monkeys), two New World species (capuchin, tamarin), and two prosimians (bushbaby, tree shrew). 2. Following intra-muscular phenol or 1-naphthol (10 mg/kg), sulphation was the major conjugation in the Old World monkeys and prosimians, whereas glucuronidation predominated in the New World species. 3. In rhesus and cynomolgus monkeys, sulphation decreased as dose increased, but remained the major conjugation with both substrates at dose levels of 0.01 to 25 mg/kg. 4. In the capuchin, the conjugation pattern of phenol changed markedly as dose increased; at 0.01 and 1 mg/kg sulphation was the major conjugation, whereas at 10 and 25 mg/kg glucuronidation predominated. With 1-naphthol only small amounts of sulphate were excreted; glucuronic acid conjugation was the major metabolism at all four dose levels. 5. The importance of considering both substrate and dose when making inter-species comparisons, particularly with man, is discussed.

Animals↗

PubMed1979

Chronologic changes of activities of naphthol AS-D acetate esterase and other nonspecific esterases in the mononuclear phagocytes of tuberculous lesions.

Nonspecific esterases of mononuclear phagocytes (MNs) were studied histochemically in the developing and healing tuberculous lesions produced in rabbit skin by bacille Calmette Guérin (BCG). Nonspecific esterases were assayed with the following substrates: naphthol AS-D acetate (AS-D), naphthol AS-D chloroacetate (AS-D Chl), naphthol AS acetate (AS) and alpha-naphthyl acetate (alpha-N), beta-Galactosidase, a lysosomal enzyme of MNs, was also assayed as a marker of MN activation. The number of MNs hydrolyzing AS-D Chl, AS, and alpha-N increased for 2 to 4 weeks after infection. These chronologic changes were similar to that of beta-galactosidase. In contrast, MNs hydrolyzing AS-D appeared predominantly in the healing lesions five to six weeks after infection. These MNs had the morphologic features of balloon-like cells. They contained few lysosomes and gathered in clumps far from the caseous center. The activity of the AS-D esterase was almost completely inhibited by various trypsin inhibitors, but not by the serine esterase inhibitor of phenylmethylsulfonyl-fluoride. These results suggest that the AS-D esterase is a trypsin-like esterase which participates in the healing of tuberculous lesions.

Animals↗

PubMed1976

Fluorescence histochemistry of peptide hormone-producing cells: observations on the nitroso-naphthol method for the demonstration of tyrosine residues.

Nitroso-naphthol reacts with tyrosine residues of peptides (and probably also proteins) to yield intensely fluorescent condensation products. This reaction forms the basis of a fluorescence histochemical procedure designed to demonstrate cells that are rich in tyrosine-containing peptides or proteins. In models the method was found to be specific for p-hydroxylated phenolic compounds. Fluorescence was induced also following formaldehyde vapour fixation. With the nitroso-naphthol technique the zymogen granules of gastric chief cells, intestinal Paneth cells, pancreatic acinar cells and certain peptide hormone-secreting cells such as the GH cells in the adenohypophysis, the insulin cells of the pancreatic islets and the calcitonin cells of the thyroid gave intense fluorescence with spectral characteristics indistinguishable from those ofthe fluorophores of tyrosine-containing peptides. In addition, a population of endocrine-like cells in the antral and intestinal mucosa of certain mammals displayed fluorescence.

Animals↗

PubMed1976

Occupational pigmented contact dermatitis from Naphthol AS.

Hyper-pigmentation as a manifestation of contact sensitivity to optical brighteners has previously been reported. In 1973 we observed an epidemic of pigmented contact dermatitis occurring in a textile mill in connection with the manufacture of drill fabrics impregnated with a recently introduced azo dye coupling component Naphthol AS. Of the 53 workers included in the study, 12 presented a spotted pattern of hyperpigmentation mainly affecting the exposed areas, but without pruritus, 18 were pigmented to a lesser degree, and in 21 it was not possible to discern any well-defined discoloration; some of the patients in the last group, however, complained of slight pruritus. The last two patients who were fair-skinned showed a reddish-brown pigmentation and a well pronounced pruritus. Hyper-pigmentation was most pronounced in individuals with dark complexions, whereas fair-skinned patient also showed "classical" eczematous symptoms including pruritus. A field study demonstrated direct exposure to the chemical. All 53 patients were patch-tested with Naphthol AS 5% in water, and 24 were found to be positive. Histological features were melanocyte proliferation, incontinence of pigment, and various degrees of damage to the basal layer.

Anilides↗

PubMed1978

Adaptation of the Naphthol Yellow S staining for objects with high protein content.

In measuring isolated rat liver cells stained with Naphthol Yellow S (NYS) at optimal conditions of pH (2.8), the absorbances measured at the absorption peak of 430 nm appeared to be far too high locally to enable accurate cytophotometric measurements. In order to bring down these absorbances, different techniques for flattening the cells, off-peak measurement and NYS staining at non-optimal pH levels have been applied respectively. Using albumin incorporated in polyacrylamide model films, the reliability of off-peak measurements and the quantitative aspects of the modified protein staining procedures have been investigated. It was found that the NYS procedure can be used as a quantitative protein staining not only at pH 2.8, but also at pH 2.0, 3.5 and 4.0 respectively. The problem with regard to the cytophotometric measuring of isolated liver cells could only be solved, however, by combining a specially developed flattening procedure (by centrifuging small drops of suspension) with staining at non-optimal pH levels. In contrast to the model film results, off-peak measurements applied in situ appeared to give rather unreliable results. In cases of a combined Feulgen-NYS staining, the Fuelgen-DNA values were not significantly influenced by any of the modifications of the original NYS staining procedure.

Animals↗

PubMed1977

Gas chromatographic determination of microamounts of carbaryl and 1-naphthol in natural water as sources of water supplies.

A method for the clean-up and quantitative determination of Carbaryl and its hydrolysis product, 1-naphthol, in natural waters is described. After extraction of a water sample with methylene chloride, the two compounds were separated from possible organochlorine pollutants such as Endrin, gamma-BHC, p,p'-DDT, pentachlorophenol and polychlorinated biphenyl, and their heptafluorobutyryl derivatives obtained. Determination by electron-capture gas chromatography at the 2.5-10 ppb level, using 11 of water samples, was carried out.

Carbaryl↗

PubMed1979

RM values of naphthols and acetophenones in structure-activity studies.

The RM values of naphthols obtained in a chromatographic system where the stationary phase consisted of a silica gel G layer impregnated with silicone oil are much more closely related to the log P values in an octanol-water system than the RM values determined on polyamide layers. Similarly, the RM values of a series of acetophenones in the silicone system are closely related to their log P values. The equations describing the structure-activity relationship indicate the importance of lipophilic character and halogen substitution in determining the hemolytic activity and the acute toxicity of compounds.

Acetophenones↗

PubMed1978

Absorption and excretion of 1-amino-2-naphthol-6-sulphonic acid in rats and rabbits.

1-amino-2-naphthol-6-sulphonic acid (ANSA) and compounds of similar structure are metabolites formed by azo reduction in mammals from several commonly used food colours, e.g. Sunset Yellow FCF and Food Red 17. At our institute it has been shown that ANSA produces bile duct proliferation when given orally to rabbits. Furthermore Orange RN which by azo reduction yields ANSA (and aniline) induce the same effect in pigs (Olsen et al., 1973) but not in rats (Gaunt et al., 1971). Using 35S-labelled ANSA the absorption and excretion as well as the localisation in the liver cell of ANSA was investigated in rats and rabbits. The results after p.o. administration shows that the rabbit absorbs and excretes into the urine a significant larger amount of ANSA (30-40%) than the rat (6-13%). The results after i.v. administration shows that some 60% of the injected dose is excreted in the urine in both species. In the rat 15-20% are recovered from the faeces, while only traces (0.1-0.5%) are found in faeces of rabbits. A significant amount of ANSA was retained in the blood and the liver of both species 24 and 48 hours after administration. After differential centrifugation of liver homogenates the majority of ANSA was found in the 9000 x g supernatant.

Absorption↗

PubMed1977

A naphthol yellow S and erythrosin B staining procedure for use in studies of the acrosome reaction of rabbit spermatozoa.

Rabbit spermatozoa suspended in Krebs-Ringer-phosphate containing 0.25% glucose were smeared on polylysine-coated slides and dried in air at room temperature for 30 min at room temperature, blotted, rinsed in 1.0% aqueous acetic acid for 10-15 sec, drained and stained for 7 min in a mixture of equal parts of aqueous naphthol yellow S and erythrosin B (final concentration of each dye 0.1% w/v) at pH 4.6-5.0 (pH adjusted with acetic acid). Stained slides were well rinsed in distilled water adjusted to pH 4.6-5.0 with acetic acid, blotted, allowed to dry completely, rinsed in xylene and mounted in synthetic resin. Acrosomal caps were stained cherry-red (apical ridge) to pink (dorsal and ventral aspects); postnuclear caps stained pale pink; nuclei were either unstained or stained a very faint yellowish-pink. The mid-piece and flagellum were stained different shades of pink. The procedure is simple, rapid, and gives highly reproducible results. When present, acrosomes are easily detected regardless of the density of the smear.

Acrosome↗

PubMed1979

Quantitative aspects of the Naphthol Yellow S staining for proteins studied in a model system of polyacrylamide films and in isolated rat liver cells and nuclei.

After staining with Naphthol Yellow S (NYS) at optimal conditions of pH (2.8), the protein content of rat liver cells isolated by means of a collagenase perfusion technique was found to be cytophotometrically immeasurable, because of too high local dye absorbances. In order to lower the absorption values, techniques to flatten the cells, off-peak measurements and NYS staining at non-optimal pH levels were applied. With polyacrylamide model films incorporated with albumin, the reliability of off-peak measurements and the quantitative aspects of the modified protein staining procedures have been investigated. It was found that NYS is a quantitative protein stain not only at pH 2.8, but also at pH 2.0, 3.5 and 4.0 respectively. Off-peak measurements can also produce quantitative results. In cases of a combined Feulgen-NYS staining, the Feulgen-DNA values were not significantly influenced by any of the NYS staining procedures.

Acrylic Resins↗

PubMed1978