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J K Chipman

Publications and source records attributed to J K Chipman.

At least 73 records · Page 4Linked to original sources
PubMed1993

DNA modification and repair by 2-nitropropane is extensive in hepatocytes of rats compared to those of humans and mice.

Hepatocytes from rats, mice or humans were exposed to either 2-nitropropane or propane 2-nitronate and the extent of unscheduled DNA synthesis (UDS) was measured. At a concentration of 2-nitropropane (0.1 mM) that produced a marked induction of UDS in rat hepatocytes, a negative or minimal response was seen in hepatocytes from mice and humans. A 10-fold higher concentration of 2-nitropropane was required in mouse hepatocytes for an equivalent UDS response to that seen in rat-liver cells. All three species showed UDS after exposure of hepatocytes to propane 2-nitronate at 0.1 mM although the effect in human cells was variable. In rat hepatocytes the induction of UDS by 2-nitropropane was not inhibited by the antioxidant dimethyl sulphoxide (1%). In these cells, 2-nitropropane produced an electrochemically active modified deoxynucleoside, similar to that reported to be formed in livers of rats which had received 2-nitropropane in vivo. A smaller but significant production of this altered deoxynucleoside was found in mouse hepatocytes but not in human hepatocytes treated identically. The level of 8-oxo-7,8-dihydrode-oxyguanosine was not increased in hepatocytes from any of the three species exposed to 2-NP under the same conditions. It is suggested that the UDS caused by 2-nitropropane may be in response to damage other than that produced by oxygen radicals.

Adult↗
PubMed1992

Bleomycin-induced unscheduled DNA synthesis in non-permeabilized human and rat hepatocytes is not paralleled by 8-oxo-7,8-dihydrodeoxyguanosine formation.

The genetic toxicity of the antitumour antibiotic bleomycin (BLM) is thought to involve the formation of a reactive oxygen intermediate. 8-Oxo-7,8-dihydrodeoxyguanosine (oxo8dG), an oxidation product of deoxyguanosine, is one of the major products formed when isolated DNA is exposed to oxygen radical generating systems. Gamma-irradiation (10-500 Gy 60Co; 10 Gy/min) or BLM and Fe2+ (37.5-150 U/L and 0.5 mM, respectively) treatment of isolated DNA (0.25 mg/mL) increased oxo8dG above background. In the latter case, the effect was greater than that with Fe2+ (0.5 mM) alone and was dependent on the dose of BLM. When DNA was irradiated with 500 Gy60Co, deoxyguanosine oxidation was inhibited by antioxidants (ethanol: 37.5 and 98% inhibition at 2 and 20 mM, respectively; mannitol: 20.5, 60 and 92% inhibition at 0.1, 1.0 and 10 mM, respectively). Similarly the BLM-induced production of oxo8dG was inhibited (64%) by mannitol (10 mM). BLM also caused production of base propenals on interaction with isolated DNA. In contrast, oxo8dG was not induced above background concentration (27 mol oxo8dG/10(6) mol dG) in permeabilized (37 degrees) and non-permeabilized (4 degrees and 37 degrees) rat hepatocytes treated with BLM (260 U/L). Despite this, there was extensive BLM-induced unscheduled DNA synthesis (10 and 100 U/L) in non-permeabilized rat and human hepatocytes in the absence of hydroxyurea. These findings, in accord with other observations, draw into question the role of .OH in BLM-induced DNA damage and the mimicry of ionizing radiation in cellular systems.

8-Hydroxy-2'-Deoxyguanosine↗
PubMed1992

Induction of 8-hydroxydeoxyguanosine in Ah-responsive mouse liver by iron and Aroclor 1254.

Treatment of Ah-responsive C57BL/10ScSn mice with iron greatly sensitizes them to induction of hepatic porphyria and tumour formation by the polychlorinated biphenyl mixture Aroclor 1254. In the present studies, male C57BL/10ScSn mice received a single dose of iron-dextran (600 mg Fe/kg) and were fed a diet containing 0.01% Aroclor 1254 for 1, 3 and 5 weeks. By use of HPLC with electrochemical detection, 8-hydroxydeoxyguanosine (8-OHdG) was then measured in liver DNA as a marker for oxidative damage. Treatments with iron or Aroclor alone did not result in a significant increase in 8-OHdG except at 3 weeks following iron treatment. At 1 and 3 weeks 8-OHdG levels were induced approximately 3- and 5-fold above control groups respectively in iron- and Aroclor-treated animals. Although there was an apparent 5- to 10-fold increase in the level of 8-OHdG at 5 weeks, this was partially attributed to the in vitro effects of porphyrins, levels of which were massively elevated in liver at this time point. The iron/Aroclor-induced synergistic elevation of 8-OHdG at 1 and 3 weeks was concluded to be due to in vivo damage, thus suggesting the importance of DNA oxidation in the early events of carcinogenesis in this system.

8-Hydroxy-2'-Deoxyguanosine↗
PubMed1992

Production of 8-hydroxydeoxyguanosine in isolated DNA by chromium(VI) and chromium(V).

Chromium(VI) and Cr(V) compounds increased the concentration of 8-hydroxydeoxyguanosine (oh8dG) in isolated DNA, whereas no such increase was seen with Cr(III). Furthermore, incubating DNA with H2O2 and Cr(VI) or Cr(V) potentiated the formation of oh8dG above levels observed with either chromium compound alone. In the presence of catalase, the increase in DNA oxidation observed with Cr(VI) was inhibited, the base oxidation observed being equivalent to background levels, and this indicated involvement of H2O2 in the mechanism. Glutathione did not enhance chromium-induced formation of this oxidized base. These results help to explain a mechanism of chromium-induced DNA oxidation involving H2O2 via a Fenton-type reaction.

8-Hydroxy-2'-Deoxyguanosine↗
PubMed1992

Induction of DNA strand breaks in peripheral lymphocytes by soluble chromium compounds.

1. Incubation of human lymphocytes with sodium dichromate (CrVI) at 37 degrees C for 3 h resulted in a dose-dependent increase in DNA strand breaks without concurrent cytotoxicity. In contrast, chromium acetate hydroxide (CrIII) failed to induce DNA strand breaks at sub-cytotoxic concentrations. 2. DNA strand breaks were also detected in the peripheral lymphocytes of Wistar rats, 24 h after intratracheal instillation of sodium dichromate (1.3 and 2.5 mg kg-1). Instillation of chromium acetate hydroxide (up to 21.8 mg kg-1) failed to induce DNA strand breaks in peripheral lymphocytes. In accord with previous studies, hexavalent chromium was found to be more readily absorbed from the lungs into the peripheral blood than chromium in its trivalent form. 3. The results of this study indicate that fluorometric analysis of DNA unwinding (FADU) in peripheral lymphocytes might be a convenient method of measuring an important biological effect of chromium in occupationally-exposed workers.

Animals↗
PubMed1991

Purification of hepatocyte couplets by centrifugal elutriation.

An initial preparation of rat hepatocytes containing approximately 30% couplets was enriched by centrifugal elutriation. Of the couplets loaded onto the elutriator, 87% were eluted at medium flow rates of 60 to 80 ml/min at a rotor speed of 1,100 rpm; cells eluted in this range maintained a viability of more than 95%. Peak fractions were enriched in couplets to 84.5% +/- 2.5%. After elutriation, couplets retained the ability to secrete fluorescent cholephiles into sealed canalicular vacuoles. The preparation can now be used in hepatobiliary and hepatotoxicity studies not possible with preparations in which they are minor components.

Animals↗
PubMed1991

Bio-techniques for the detection of genetic toxicity in the aquatic environment.

This article reviews the application of sensitive biotechniques in the detection of DNA damage within the aquatic environment as "early warning" indicators of pollution-related genetic toxicity. Bacterial mutagenicity assays have played an important role both in the discovery of mutagens and in the analysis of the ability of aquatic organisms to convert inert substances to reactive genotoxic products. Genetic damage in aquatic species treated with carcinogens has been detected by analysis of DNA adducts, induction of DNA repair, DNA strand breakage and formation of chromosomal aberrations and sister chromatid exchanges. Little information is available on pollution-related DNA damage resulting from field exposure. A 32P-postlabelling technique for the sensitive detection of DNA adducts is a promising development towards this aim and the analysis of changes in oncogene nucleotide sequence in tumour tissue is considered an important future direction.

Animals↗
PubMed1991

The measurement of DNA strand breaks in rat colonic mucosa by fluorometric analysis of DNA unwinding.

We describe the detection of DNA strand breaks in the rat colonic mucosa (in vivo and in vitro) by fluorometric analysis of DNA unwinding. DNA strand breaks were detected, in a dose-dependent manner, 2 h following an intracaecal dose of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG, 5-100 mg/kg). Levels of double-stranded DNA in the colonic mucosa of control animals, after 60 min unwinding time, were 62.8 +/- 3.4% (n = 6) and were significantly reduced at 5 mg/kg to 52.1 +/- 1.9% (n = 3). DNA strand breaks were also detected in isolated rat colonic mucosal cells following 15 min treatment with MNNG (0.1-1 mM) in vitro. The percentage of double-stranded DNA, after 60 min unwinding time, was significantly reduced (13.6 +/- 5.4%, n = 3) from control levels following treatment with 0.1 mM. A post-treatment incubation (30 min) of cells treated at a MNNG concentration of 0.25 mM resulted in 78.2 +/- 24.0% (n = 3) of these lesions being repaired.

Animals↗
PubMed1991

Lack of release from hepatocytes in vitro or excretion in vivo of mutagenic chrysoidine metabolites.

Rat liver postmitochondrial supernatant (S9) converted the azo dyes chrysoidine Y and R to products that were mutagenic towards Salmonella typhimurium strain TA100. No such release of mutagens was demonstrated using intact rat hepatocytes as an activation system despite the fact that chrysoidine dyes cause unscheduled DNA synthesis in these cells. It appears that genotoxic products produced within hepatocytes either react within the cell or are detoxified prior to release. Following intraperitoneal administration of chrysoidine Y to rats (100 mg/kg i.p.) there was also no evidence of mutagenic or por-mutagenic products excreted in bile or urine. The S9-derived mutagens appear to be largely independent of bacterial acetylation since they were active in the acetylation-deficient strain TA98/1,8-DNP6 in addition to strain TA98. The ultimate mutagenic form(s) are therefore unlikely to be acetoxyarylamines.

Animals↗
PubMed1990

Bacterial mutagenesis and hepatocyte unscheduled DNA synthesis induced by chrysoidine azo-dye components.

5 azo dye components of Gurr chrysoidine 'Y' have been separated, synthesised and identified. Dyes with a methyl substitution (particularly between the two amino groups) were more mutagenic in Salmonella typhimurium strain TA100 with control rat liver S9 than the non-methylated counterpart (range 66-1992 revertants at 50 micrograms/plate). Mutagenicity was also catalysed by human-liver S9 and pre-treatment of rats with either phenobarbitone or beta-naphthoflavone enhanced the activation ability of S9 by greater than 4-fold. Using the most potent promutagenic component (2,4-diamino-3-methylazobenzene), the use of inhibitors of cytochrome P450 (metyrapone: 1.0 mM; alpha-naphthoflavone: 0.075 mM; DPEA: 0.125 mM) and of the flavin monooxygenase (methimazole: 0.75 mM) suggested a major role for cytochrome P448 in the activation of chrysoidine to mutagens. The ability of chrysoidine components to induce unscheduled DNA synthesis in rat hepatocytes in vitro was demonstrated and ranged between 11.92 and 23.5 net nuclear grains at a dose level of 2.5 micrograms/incubation. Since each dye was equi-potent, methyl substitution had little influence on genetic toxicity in hepatocytes.

Animals↗
PubMed1990

Direct modulatory effect of hexasodium N,N,N',N'-ethylenediamine-tetramethylene-phosphonate on bone cell function in vitro.

The phosphonate hexasodium N,N,N',N'-ethylenediamine-tetramethylenephosphonate (EDITEMP.Na6) reduced alkaline phosphatase (Alp) activity and cAMP response to parathyroid hormone (PTH) in primary cultures of foetal rat calvaria cells in a dose-dependent manner, while not affecting culture DNA content. EDITEMP.Na6 also inhibited the mineralization of three-dimensional bone nodules formed in vitro, but not the number of nodules formed. Bone cell culture DNA content was also reduced by EDITEMP.Na6 but at concentrations in excess of those needed to modulate osteoblastic cell function. Withdrawal of EDITEMP.Na6 led to slow but complete recovery of Alp activity. At EDITEMP.Na6 concentrations of 25 microM and higher, recovery of Alp activity appeared to be independent of protein and/or DNA synthesis. Cell culture acid phosphatase (Acp) activity was not affected by EDITEMP.Na6. The results indicate that EDITEMP.Na6 has a direct inhibitory effect on (mature) osteoblastic cell function. In the presence of bone tissue this inhibition also occurred, although not at a relatively low dose level.

Acid Phosphatase↗
PubMed1990

Effects of intraperitoneal administration of hexasodium N,N,N',N'-ethylenediaminetetramethylenephosphonate on rat bone metabolism.

The influence of hexasodium N,N,N',N'-ethylenediaminetetramethylenephosphonate (EDITEMP.Na6) on biochemical parameters in bone has been studied to help explain recently reported histological and pathological changes in bone induced by this agent. A single, intraperitoneal (ip) injection of EDITEMP.Na6 (350 mumol/kg on Day 0) significantly decreased the activity of serum alkaline phosphatase (Alp) in mature male Wistar rats for up to 10 days following the injection. However, in immature rats the initial EDITEMP.Na6-induced reduction of serum Alp activity preceded a steady increase in serum Alp activity, which was significantly elevated between Days 6 and 13. This increase coincided with a significant increase in bone Alp activity (an indicator of bone formation) on Day 8. Diaphyseal acid phosphatase (Acp) activity (an indicator of bone resorption) was significantly reduced, on Days 4 and 8 but not on Day 1, following treatment with EDITEMP.Na6. Repeated exposure of immature male Wistar rats to EDITEMP.Na6 (70 mumol/kg/day, ip for 7 days) also led to a significant and persistent increase in serum and bone Alp activities and a reduction in diaphyseal Acp activity. The data are in accord with published histomorphological evidence of reduced bone resorption and suggest delayed mineralization of osteoid tissue laid down during the treatment with EDITEMP.Na6. There were only transient and limited changes in bone DNA content and [3H]thymidine incorporation throughout the studies, indicating the absence of marked cytotoxicity or altered bone cell proliferation associated with EDITEMP.Na6-induced changes in phosphatase enzyme activities.

Acid Phosphatase↗
PubMed1989

Formation of nuclear anomalies in rat intestine by benzidine and its biliary metabolites.

Administration of benzidine (BZ) by intraperitoneal injection (i.p.) to rats (0-100 mg/kg) produced, after 24 h, a dose-dependent formation of nuclear anomalies (micronuclei, pyknotic and karyorrhectic nuclei) in intestinal epithelial cells analysed both in isolated cell suspensions and in the intestinal crypts in tissue sections. When bile collected (0-4 h) from rats treated with BZ (150 mg/kg, i.p.) was infused into the duodenum of recipient rats, nuclear anomalies were observed in mucosal epithelial cells, after 24 h, with a similar distribution to that in rats given BZ by i.p. injection. The formation of nuclear anomalies in the intestine is in accord with the intestinal carcinogenic effect of BZ and is, at least partially, dependent on exposure of epithelial cells to biliary metabolites of BZ.

Animals↗
PubMed1989

Comparison of the hepatotoxicity in mice and the mutagenicity of three nitroalkanes.

The hepatotoxic and mutagenic potentials of 2-nitropropane, nitromethane, and nitroethane were compared. Hepatotoxicity was assessed biochemically and histopathologically in BALB/c mice. In male mice, plasma activities of the hepatic enzymes sorbitol dehydrogenase, alanine aminotransferase, and aspartate aminotransferase were significantly elevated 48, 72, and 96 hr after ip administration of 9 mmol/kg 2-nitropropane, but not at 24 hr and not after administration of smaller doses of 2-nitropropane nor after nitromethane or nitroethane (9 mmol/kg). In female mice a dose of 6.7 mmol/kg of 2-nitropropane was sufficient to cause hepatotoxicity. The histopathological evaluation supported the biochemical results, and livers of mice that had received 2-nitropropane (9 mmol/kg) showed damage, particularly in the periportal region. Mutagenicity was tested in Salmonella typhimurium tester strains TA98, TA100, and TA102. Both 2-nitropropane and its anionic form, propane-2-nitronate, were mutagenic but the nitronate was the more powerful mutagen. Nitromethane, nitroethane, nor their nitronates caused an increase in the number of revertant colonies over those seen in control plates. The results suggest that the primary nitroalkanes are much less hepatotoxic and mutagenic than 2-nitropropane.

Animals↗
PubMed1989

The use of flow cytometry in the detection of radiation- and chemically induced nuclear damage in human mononuclear cells.

Viable human mononuclear cells were treated with three known genotoxic agents, gamma radiation, bleomycin sulphate (BLM) and acetylaminofluorene (AAF). Following treatment, a high salt concentration buffer was used to disrupt the cells, releasing supercoiled DNA structures known as nucleoids. The nucleoids were treated with the fluorescent dye ethidium bromide, and exposed to a beam of laser light within a fluorescence activated cell sorter (FACS). Treatment of human mononuclear cells with the genotoxic agent BLM or with radiation prior to cell disruption and staining produced a dose-dependent increase in light scatter. However, treatment of human mononuclear cells with AAF, which requires metabolic activation, produced a dose response in light scatter that proved reversible and was not dependent on the formation of reactive metabolites. In the case of BLM and radiation, these changes in light scatter are interpreted as structural alterations in the nucleus partially induced by free radical attack, and thus provide an indication of the DNA damaging ability of the agent used. In contrast the reversible changes in light scatter seen with AAF may only indicate the ability of this agent to penetrate the genome. The significance of this reversible interaction with respect to genotoxicity is not known.

2-Acetylaminofluorene↗
PubMed1989

Host-mediated bacterial mutagenesis and enterohepatic circulation of benzidine-derived mutagenic metabolites in rodents.

1. Administration of benzidine (100 mg/kg, i.p.) to bile duct-cannulated rats led to a sustained excretion of metabolites in bile which, following glucuronide hydrolysis, were mutagenic to Salmonella typhimurium strain TA98. 2. When the biliary metabolites were re-infused into the duodena of a further group of rats, enterohepatic circulation of mutagens was indicated by extensive re-excretion of biliary mutagens in the recipients. 3. Furthermore, in mouse host-mediated mutagenicity assays, both i.p. injection of benzidine (100 mg/kg) and intracaecal administration of rat biliary metabolites of benzidine produced a mutagenic response in Salmonella typhimurium strain TA98 cells isolated from the liver. 4. The results indicate that enterohepatic circulation adds to the biological persistence of reactive metabolites of benzidine and may contribute to the carcinogenicity of this aromatic amine.

Animals↗