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M Marion

Publications and source records attributed to M Marion.

At least 19 recordsLinked to original sources
PubMed1992

Effect of 2-acetylaminofluorene on intracellular free Ca2+ in isolated rat hepatocytes.

The effect of 2-acetylaminofluorene (2-AAF) on the intracellular free Ca2+ ([Ca2+]i) and viability of isolated rat hepatocytes has been investigated using the fluorescent probes quin 2 and propidium iodide respectively. At the highest concentration tested (224 microM), 2-AAF produces an elevation of [Ca2+]i which shows a biphasic profile. A small initial increase is observed during the first 5 min; this is followed by a considerable rise which reaches up to 2.5 times the control value at 15 min. These changes in intracellular calcium are not accompanied by detectable alterations in cell viability. In order to determine the mechanisms by which this effect of 2-AAF takes place, three calcium antagonists, namely verapamil, TMB-8 (8-(diethylamino)-octyl-3,4,5-trimethoxybenzoate) and ruthenium red (RuR), have been used. The results suggest that the first phase is dependent upon internal Ca2+ store mobilization, while the second phase seems to be related to Ca2+ entry from the extracellular space. The data obtained with RuR further indicate that mitochondria may be involved in the perturbation of calcium homeostasis caused by 2-AAF. In addition, in the experiments involving antagonists, no consistent pattern emerges that suggests a close relationship between intracellular Ca2+ levels and cell viability. The present study provides further information on the mechanisms by which these well-known hepatotoxin 2-AAF may interact with liver cells. It also shows that when these cells are exposed to a toxin, short-term changes in [Ca2+]i may not be accompanied by loss of cell viability, and conversely, that changes in cell viability may occur without alterations in [Ca2+]i.

2-Acetylaminofluorene↗
PubMed1992

Computerised infrared thermography and isotopic bone scanning in tennis elbow.

Thirty five cases of tennis elbow (17 unilateral, nine bilateral) were studied with infrared thermography and isotopic bone scanning. A hot focus was visualised in 16 of 17 cases of unilateral tennis elbow (94%) and in all nine cases of bilateral tennis elbow (100%) on infrared thermography, and abnormal increased epicondylar activity seen in 12 of 17 (71%) and eight of 18 (44%) cases respectively with isotopic bone scanning. Unilateral visual cooling (somatosympathetic responses) occurred in seven of 13 cases of unilateral tennis elbow (54%) with infrared thermography, and reduced perfusion in seven of 12 (58%) of similar cases with blood pool isotopic bone scanning. Computerised temperature assessments showed statistically significant side to side temperature differences when 17 active tennis elbows were compared with the opposite normal elbows for spot temperatures, proximal and distal forearm gradients. Similar temperature assessments in 18 bilateral tennis elbows compared with 17 normal elbows showed significant temperature differences for elbow spot temperatures and distal forearm gradients, but not for proximal gradients.

Adult↗
PubMed1990

The cytotoxic effects of cadmium chloride and mercuric chloride mixtures in rat primary hepatocyte cultures.

The toxic effects of Cd and Hg mixtures were studied using primary monolayer cultures of rat hepatocytes. Cytotoxicity was assessed by measuring the release of lactic dehydrogenase from the cells. Cytotoxic and non-cytotoxic metal levels were used. At the higher exposure concentrations (0.2 micrograms Cd.ml-1 and 2.0 micrograms Hg.ml-1), Cd was very toxic to hepatocytes whereas Hg was only marginally toxic. The combination of Cd and Hg was more toxic than predicted by summation of the individual metal toxicities. The incorporation of [35S]cysteine into protein of the cytosol and insoluble cell fraction was increased in response to Cd or Hg exposure and was directly related to cell 35S accumulation. Combinations of Cd and Hg significantly increased the proportion of total 35S which was incorporated in cell protein, an effect that was attributed to the accumulation of protein in the insoluble cell fraction. Cd uptake by hepatocytes was related to exposure concentration but was lower when Hg was also present in the incubation medium. Gel chromatography of the cytosol from Cd-exposed cells showed 3 Cd containing fractions which corresponded to the elution positions of high Mr proteins, metallothionein (MT) and low Mr molecules. When hepatocytes were exposed to Hg in combination with Cd, the MT-like fraction was no longer evident and Cd in the low Mr fraction was greatly reduced. Regardless of the presence or absence of Cd in the exposure medium, 98% of cytosol Hg in Hg-exposed cells was found to elute after the low Mr fraction, at a position equivalent to inorganic salts. This indicates that the enhanced cytotoxicity of Cd and Hg may be related to a decrease in the MT-like protein in the cytosol and not due to a direct competitive binding interaction in relation to the protein.

Animals↗
PubMed1990

Metallothionein induction and metal homeostasis in rainbow trout hepatocytes exposed to mercury.

Rainbow trout hepatocytes were exposed to a sublethal concentration of mercury (Hg) (100 nM) for 0, 5, 10, 15, 20, 25, 30 and 35 h at 15 degrees C. The hepatocytes were found to accumulate appreciable quantities of Hg. The presence of Hg led to elevated Ca, Cu and Zn levels in cells. Hg was bound mainly to low-molecular-weight components, although it induced the synthesis of metallothionein (MT). The results suggest that when hepatocytes are exposed to low levels of Hg, MT does not play a significant role in the sequestration of the metal. The elevated cellular Cu and Zn levels could be associated with the induction of MT.

Animals↗
PubMed1990

Metal homeostasis and metallothionein induction in rainbow trout hepatocytes exposed to cadmium.

This study was undertaken to investigate the cellular effects of Cd and metallothionein (MT) metabolism using a primary cell culture system composed of isolated rainbow trout (Salmo gairdneri) hepatocytes. The cells were exposed to 89 nM Cd for 0, 5, 12, 20, 25, 30, 35, and 40 hr at 15 degrees C. The concentration of Cd used did not cause significant cell damage, as estimated by lactate dehydrogenase (LDH) release into the extracellular medium. However, the presence of the metal led to increased cellular levels of calcium within the first 12 hr of incubation. Later, these returned to control values and remained as such for the rest of the examination period. A transient increase in intracellular copper and zinc also occurred during the initial incubation period. In parallel, the hepatocytes were found to accumulate appreciable quantities of Cd, a significant proportion of which were detected in the cytosol. Distribution profiles of cytosolic Cd obtained through Sephadex G-75 chromatography showed that the metal was associated mainly with the high-molecular-weight (HMW) and middle-molecular-weight (MMW) protein fractions. At t = 12 hr, there was a maximum in the proportion of Cd in the MMW fractions, which was accompanied by a decrease in the proportion of Cu in the same fractions. At this time, cellular MT exhibited the highest levels. MMW fractions were further resolved using anion-exchange chromatography. Although Cd was present in the peaks corresponding to MT, the data indicated that these peaks also contained detectable amounts of Cu and Zn.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Genotoxic effects of heavy metals in rat hepatocytes.

The genotoxic interaction of metals, which are common environmental contaminants, was studied in cultured hepatocytes. Freshly isolated rat hepatocytes were exposed to concentrations of cadmium, copper, silver and lead salts ranging from non-cytotoxic to moderately cytotoxic (as determined by LDH release), and the incorporation of [3H]thymidine into the DNA, as a measure of repair synthesis, was followed. In addition, the uptake of metals by the nuclear fraction was determined using Inductively Coupled Plasma/Mass Spectrometry or atomic absorption spectrophotometry. The evaluation of binding of 109Cd to the DNA in situ was also attempted. It was observed that after a 20 h exposure period, all the metals investigated were found in the nuclear fraction of hepatocytes, with Ag apparently being accumulated less efficiently. In parallel, Cd (0.18 to 1.8 microM) and Cu (7.9 to 78.5 microM) consistently produced a statistically significant stimulation of [3H]thymidine incorporation into the DNA, in the presence or absence of hydroxyurea while Ag was active only at the highest concentration tested (18.5 microM). In contrast, Pb failed to induce a UDS response at the levels used. Moreover, exposure of hepatocytes to 1.8 microM 109CdCl2 for 20 h led to a DNA binding ratio of 0.98 +/- 0.23 ng Cd/micrograms DNA. The present results support the view that the nucleus may be an important target organelle for metal toxicity.

Animals↗
PubMed1989

The effects of putative DNA repair inhibitors on DNA adduct levels and unscheduled DNA synthesis in rat hepatocytes exposed to 2-acetylaminofluorene.

The enzymology of DNA repair is currently under active investigation. The purpose of the present study was to examine the involvement of a number of enzymes (DNA polymerase alpha and beta, DNA topoisomerase II and ribonucleotide reductase) in the repair of chemically induced DNA damage in a mammalian cell system. This was done by studying the effects of inhibitors of these enzymes on the levels of 2-acetylaminofluorene (2-AAF)-DNA adducts and on the induction of UDS in primary cultures of rat hepatocytes exposed to the carcinogen in vitro. The results obtained with aphidicolin (an inhibitor of DNA polymerase alpha) show that the binding of 2-AAF to cellular DNA was significantly higher in samples exposed to this compound. Moreover, induction of UDS by 2-AAF was completely blocked in the presence of this compound. Dideoxythymidine, a DNA polymerase beta inhibitor, led to complex results. It produced a reduced DNA-specific activity due to [3H]2-AAF adduct formation as well as a diminished but still detectable UDS response in the presence of 2-AAF. Inhibitors of DNA topoisomerase II (nalidixic acid) and ribonucleotide reductase (hydroxyurea) did not cause any statistically significant change in the accumulation of 2-AAF adducts nor did they affect the induction of UDS. The data clearly suggest that DNA polymerase alpha participates in the repair of 2-AAF adducts in hepatocytes. In addition, neither DNA topoisomerase II activity, nor limitations in the precursor nucleotide pools appear to be critical factors in this process.

2-Acetylaminofluorene↗
PubMed1989

Inhibition of binding of 2-acetylaminofluorene to DNA by butylated hydroxytoluene and butylated hydroxyanisole in vitro.

Numerous studies have shown that the food antioxidants butylated hydroxytoluene (BHT) and butylated hydroxyanisole (BHA), under specific exposure conditions, can inhibit hepatocarcinogenesis induced by various carcinogens. The purpose of the present work was to study the biochemical mechanisms responsible for the anticarcinogenic activity of BHA and BHT using in vitro systems. The effects of BHA and BHT on the binding of 2-acetylaminofluorene (2-AAF) to DNA was determined in a microsomal system and in primary cultures of rat hepatocytes. It was found that both antioxidants reduce the binding of 2-AAF and that of N-OH-2-acetylaminofluorene (N-OH-2-AAF) to calf thymus DNA in the presence of liver microsomes. The inhibition was however more pronounced with the parent compound. Lower levels of DNA binding were also detected in hepatocytes incubated with 2-AAF along with BHA or BHT. These results suggest that phenolic antioxidants can exert anticarcinogenic activity through modulation of carcinogen interaction with DNA which may reflect on alteration in carcinogen metabolic activation.

2-Acetylaminofluorene↗
PubMed1987

The modulation by metallothionein of cadmium-induced cytotoxicity in primary hepatocyte cultures.

The cytotoxicity of CdCl2 and 2 isoforms of hepatic cadmium-metallothionein (CdMT I and II), was investigated using primary cultures of rat hepatocytes. The cell cultures were exposed to cadmium as CdCl2 or as either isoform of CdMT for a 20-h period at concentrations ranging from 50 to 500 ng Cd X ml-1. Cytotoxicity was assessed by determining the amount of lactic dehydrogenase released from the cells into the incubation medium and the incorporation of [3H] arginine into cell protein. The uptake of Cd by the cells was also measured. Cadmium chloride and both isoforms of CdMT were found to be toxic to hepatocytes although partial protection was afforded by the binding of cadmium to metallothionein (MT). At the higher exposure concentrations and in accordance with the toxicity data, the cells exposed to CdCl2 were found to accumulate more cadmium than those exposed to CdMT. The distribution of cadmium in the culture medium was examined using Sephadex G-75 chromatography. The speciation of cadmium is discussed in relation to its cytotoxicity.

Animals↗
PubMed1985

Absence of genotoxic effects of nonasbestos mineral fibers.

The biological activity of natural and synthetic mineral fibers has been examined. Natural attapulgite [(Mg, Al)2Si4O10(OH).4H2O], synthetic xonotlite [Ca3Si3O8(OH)2] and natural sepiolite [Mg2Si3O8.2H2O] were selected. Genotoxic effects were investigated by means of a well established cellular model based upon the measurement of unscheduled DNA synthesis (UDS) in rat hepatocytes in primary culture. The intrinsic capacity of the fibers (1 and 10 micrograms/ml) to induce UDS was first tested. None of the fiber types showed detectable UDS-eliciting activity. Also, the possible modulation of the cellular response to genotoxic agents by the materials was examined by exposing the cells to mixtures of 2-acetylaminofluorene (AAF) (0.05 and 0.25 micrograms/ml) and fibers (1 and 10 micrograms/ml). In these experiments, the UDS response was significantly diminished in the presence of xonotlite. This phenomenon may reflect changes in the uptake and/or metabolism of AAF or may result from an inhibition of DNA repair processes, the latter suggesting a possible cocarcinogenic potential for this synthetic silicate. These results point to the immediate necessity of studying more extensively the biological effects of fibrous materials that can be used as substitutes for asbestos.

Animals↗
PubMed1985

Ultrastructural study of mineral fiber uptake by hepatocytes in vitro.

Cellular interactions of a series of fibrous materials were examined by the use of a well established in vitro system. Primary cultures of hepatocytes were exposed to natural attapulgite, synthetic xonotlite and natural sepiolite. Ultrastructural analyses revealed that hepatocytes can engage in the phagocytosis of all 3 types of fibers over an exposure period of 20 h. Attapulgite fibers were found in plasma membrane invaginations, and deeper in the cytoplasm, in vesicles exhibiting various shapes. Xonotlite was also incorporated in plasma membrane invaginations; furthermore, these fibers were present in large vacuoles where they were circumscribed by membranes and appeared somewhat isolated from the cytoplasm. Sepiolite fibers were also taken up by the cells and could likewise be identified in the previously described structures. These observations point to the relevance of the hepatocyte model for investigating the effects of fibrous materials at the cellular level.

Animals↗
PubMed1985

Genotoxicity of dimethylnitrosamine in the presence of chrysotile asbestos UICC B and xonotlite.

Interactions of particulates with chemical genotoxic agents may play an important role in the induction of carcinogenesis. With respect to bronchogenic cancer, the synergism associated with combined exposure to asbestos and tobacco smoke is a well-documented phenomenon. The present work focused on chrysotile asbestos and xonotlite. The latter is a fibrous calcium silicate which is increasingly being used to replace asbestos in various industrial applications. The study was aimed at testing the possible interaction of these materials with dimethylnitrosamine (DMN), a genotoxic component of tobacco smoke. The capacity of fibers to interfere with the genotoxic response elicited by DMN in the UDS/hepatocyte assay system specifically designed for sensitive detection of short-patch DNA repair, was looked for. The properties of the selected fibers with respect to binding affinity towards DMN were also examined.

2-Acetylaminofluorene↗
PubMed1985

Temporal bone fractures: a histopathologic study.

Trauma or injury to the skull, if of sufficient force, may produce a temporal bone fracture. However, terms such as basilar skull fracture, labyrinthine hemorrhage, or labyrinthine concussion are inadequate in that they do not describe the precise histopathologic lesion. We carried out a histopathologic study of the 14 ears in seven patients; all the patients suffered at least one temporal bone fracture. Our objective was to review the histopathologic findings of various fracture lines through the base of the skull and correlate these with the history and clinical findings.

Adult↗
PubMed1985

Persistence of the stapedial artery: a histopathologic study.

Persistence of the stapedial artery is a rare event. Fewer than 30 cases have been reported since the discovery of this artery in 1836. We carried out a histopathologic study on three temporal bones from two patients who had this anomaly, and were able to trace the full course of the artery. In two specimens a large stapedial artery persisted and substituted for the middle meningeal artery. In the third, a small, persistent stapedial artery ended in the arterial plexus surrounding the facial nerve. Persistence is discussed in terms of embryogenesis, developmental theories, histologic findings, and clinical significance. The material suggests that the stapedial artery can persist to varying degrees.

Arteries↗
PubMed1979

Mode of action of carbimazole in Graves' disease.

In a series of hyperthyroid subjects treated with a high dose 'block-replace' regime, mean thyroidal technetium uptake was shown to fall progressively over an 18 month period. Furthermore, sub-total inhibition of hormone biosynthesis could be demonstrated, without evidence of proportionate inhibition of organification, the regression of radio-iodine on technetium thyroid uptake showing similar slopes in euthyroid, treated and untreated hyperthyroid subjects. Together with the reported differential in remission rates between propranolol and anti-thyroid drugs, it is concluded that carbimazole does not act solely by inhibiting intrathyroid hormono-genesis, but more definitively by affecting thyroid hyperstimulation at a pre-biosynthetic level. The inability to demonstrate normal suppressibility of trapping in any subject in the present series despite extended therapy with a 'block-replace' regime, further suggests that current concepts of 'suppressibility' are invalid, the progressive fall in technetium uptake being unlikely to represent spontaneous restoration of a normal pituitary-thyroid axis, but rather a direct influence or prolonged therapy with anti-thyroid drugs on the natural history of the disease.

Adolescent↗