ABSTRACT
OBJECTIVE: This study tested the hypothesis that successful periodontal treatment was associated with a reduction in the incidence of spontaneous preterm birth (PTB). DESIGN: This was a randomised, controlled, blinded clinical trial. SETTING: Hospital outpatient clinic. POPULATION: Pregnant women of 6-20 weeks of gestation were eligible. METHODS: Of 322 pregnant women with periodontal disease, 160 were randomly assigned to receive scaling and root planing (SRP, cleaning above and below the gum line), plus oral hygiene instruction, whereas the remaining 162 received only oral hygiene instruction and served as an untreated control group. Subjects received periodontal examinations before and 20 weeks after SRP, and were classified blindly according to the results of treatment into two groups: successful ('non-exposure') and unsuccessful ('exposure') treatment. Groups were compared using standard inferential statistics; dichotomous variables were compared using the chi-square test or logistic regression. Results are presented in terms of odds ratios. MAIN OUTCOME MEASURE: The main outcome measure was spontaneous preterm birth before 35 weeks of gestation. RESULTS: No significant difference was found between the incidence of PTB in the control group (52.4%; n = 162) and the periodontal treatment group (45.6%; n = 160) (P < 0.13, Fisher's exact test). The incidence of PTB was compared within the periodontal treatment group, considering the success of therapy. A logistic regression analysis showed a strong and significant relationship between successful periodontal treatment and full-term birth (adjusted odds ratio 6.02; 95% CI 2.57-14.03). Subjects refractory to periodontal treatment were significantly more likely to have PTB. CONCLUSIONS: A beneficial effect on PTB may be dependent on the success of periodontal treatment.
Subject(s)
Bacterial Infections/complications , Dental Scaling/methods , Oral Hygiene/methods , Periodontal Diseases/complications , Premature Birth/microbiology , Adult , Double-Blind Method , Female , Humans , Periodontal Diseases/therapy , Pregnancy , Pregnancy Complications, Infectious , Pregnancy Outcome , Prenatal Care/methods , Young AdultABSTRACT
Grapes in Marlborough are typically grown on a vertical shoot positioned trellis system (VSP). For this purpose Pinus radiata posts are treated with CCA, a mixture of copper (Cu), chromium (Cr) and arsenic (As), giving a wood concentration of 1,730, 3,020 and 2,410 mg/kg, respectively on a dry matter basis. The CCA levels around the posts in different soils were investigated and assessed for the potential leaching of CCA into ground water. An initial survey showed leaching of all three heavy metals from the treated posts into the soil surrounding the posts (0.2% of the total vineyard area) compared with the control, depending on vineyard age and soil type. The rate of movement out of the posts was calculated from posts placed in lysimeters. HortResearch's Soil Plant Atmosphere Model (SPASMO) was used to predict the leaching rate of CCA. For As, leaching was found to be 5 mg/post/month, with the Cr rate being about twice that. Further modelling revealed a steady plume of As moving downwards after about 200-300 years. However, long-term hydrogeological modelling showed that sufficient aquifer water flow prevented the accumulation of CCA in the ground water. The modelling approaches are discussed.
Subject(s)
Water Pollution , Wine , Arsenic/analysis , Chromium/analysis , Copper/analysis , Kinetics , Models, TheoreticalABSTRACT
Despite the many advances in medicine, the rate of preterm birth has not significantly decreased in the United States over the past several decades. In fact, the rate rose in 2003 to more than 12% of all births in the United States. This equates to over half a million premature births in the United States alone. Consequently, the identification of risk factors for preterm birth which are amenable to intervention would have far-reaching and long-lasting effects. There is emerging evidence of a relationship between periodontal health and adverse pregnancy outcomes, particularly preterm birth/preterm low-birth-weight infants. Therefore this chapter explores the putative association between periodontal disease and infant prematurity, as well as the results of intervention studies which treated periodontal disease in order to reduce the incidence of prematurity. Of 31 published studies, 22 show a positive association between premature birth and periodontal disease. Ongoing studies are addressing the efficacy of periodontal treatment for decreasing the incidence of infant prematurity.
Subject(s)
Periodontal Diseases/etiology , Periodontal Diseases/therapy , Pregnancy Complications/etiology , Pregnancy Complications/therapy , Female , Humans , Periodontal Diseases/diagnosis , Pregnancy , Pregnancy Complications/diagnosis , Pregnancy Outcome , Premature Birth/etiology , Risk FactorsABSTRACT
Boron (B) is a widespread environmental contaminant that is mobile relative to other trace elements. We investigated the potential of hybrid poplar (Populus sp.) for B phytomanagement using a lysimeter experiment and a field trial on B-contaminated wood-waste. In both studies, poplars enhanced evapotranspiration from the wood-waste, reduced B leaching, and accumulated B in the aerial portions of the tree. When grown in a substrate containing 30 mg/kg B, poplar leaves had an average B concentration of 845 mg/kg, while the stems contained 21 mg/kg B. Leaf B concentrations increased linearly with leaf age. A decomposition experiment revealed that abscised leaves released 14% of their B during the winter months. Fertiliser application enhanced tree growth without decreasing the leaf B concentrations. Harvesting alternate rows of trees on a contaminated site would reduce leaching from the site while removing B. Harvested plant material may provide bioenergy, stock fodder, or an amendment for B-deficient soils.
Subject(s)
Boron/toxicity , Industrial Waste , Populus/physiology , Soil Pollutants/toxicity , Wood , Biodegradation, Environmental , Boron/analysis , New Zealand , Plant Leaves/chemistry , Soil Pollutants/analysisABSTRACT
With a fast change of land use in Marlborough from extensive pastoral farming to intensive irrigated viticulture, a need has risen to investigate the sustainable use of the available water. In 2001 a 5 ha irrigation research project was installed in a Marlborough Sauvignon Blanc vineyard. Irrigation treatments installed were control (compensate 100% for crop evapotranspiration (ET(O)), 80%, 70% and 60% of ET(O). During the two years that the Regulated Deficit Irrigation (RDI) trial has run so far, very different climatic conditions created much greater differences in yield and vegetative growth, than up to 40% reduction in irrigation, none of which were significant. The use of sap flow in the vines has been fine-tuned and is now giving reliable results on which to base vine water need.
Subject(s)
Conservation of Natural Resources , Vitis/growth & development , Water Supply/standards , Agriculture , Climate , Environmental Monitoring , New Zealand , Vitis/physiologyABSTRACT
Allophanic soils are widespread around the world, but little research has been done on their transport properties. This study reveals the effect of two soil water potential heads and two water-flow regimes of continuous and intermittent flow on solute transport through undisturbed soil columns of Horotiu silt loam (Typic Hapludand), an allophanic soil. Two different methods--breakthrough curves (BTCs) and time domain reflectometry (TDR)--were employed to determine the extent of preferential solute transport in the topsoil. The TDR data were also used to look at the depth dependence of the transport properties. The convection-dispersion equation (CDE) with the appropriate boundary conditions adequately described the movement of both Br and Cl under the various flow conditions. Although no preferential flow was found under the imposed unsaturated flow conditions, the flow of water and transport of solute became more uniform with depth. The results show that both Br and Cl are retarded in this allophanic soil. Retardation values range from 1.5 to 1.9, and, as the TDR data showed, increase from the depth of 5.0 to 10.0 cm. Intermittent leaching results showed that there was no effect on solute concentrations in the leachate following no-flow periods. This suggests that water and solute transport in this soil were either relatively uniform or that transverse mixing during flow was already fast enough to eliminate concentration gradients between regions of different "mobility."
Subject(s)
Bromides/analysis , Chlorides/analysis , Soil Pollutants/analysis , Water Pollutants, Chemical/analysis , Humans , Models, Theoretical , Soil/analysis , Volcanic Eruptions/analysis , Water MovementsABSTRACT
Land treatment of dairy-farm effluent is being widely adopted as an alternative to disposal into surface waters in New Zealand. This study investigated water balances and associated N leaching from short-rotation forest (SRF) species irrigated with dairy-farm effluent. Single trees were grown in lysimeters filled with Manawatu fine sandy loam (mixed mesic Dystric Eutrochrept). Dairy-farm effluent was applied during two irrigation periods at 21.5 mm wk(-1) with a total loading equivalent to 870 kg N ha(-1) occurring over 17 mo. Following tree harvest in April 1997, measurements continued until August 1997 to monitor tree reestablishment. Cumulative N leached did not differ between lysimeters in which evergreen Sydney blue gum (Eucalyptus saligna Sm.) and shining gum [Eucalyptus nitens (H. Deane & Maiden) Maiden] and deciduous kinu-yanagi (Salix kinuyanagi Kimura) were grown. Leachate N concentrations of all treatments were on average higher than the New Zealand drinking water standard of 11.3 mg N L(-1). The E. nitens and S. kinuyanagi treatments leached 33 and 35 kg N ha(-1) yr(-1) in 1996 following application of 236 kg N ha(-1) during the first irrigation season. Leaf area was strongly correlated to evapotranspiration, drainage volume, and nitrogen leached. The majority of leaching in the tree treatments occurred after harvest. Reducing the leaching in the regrowth phase may be achieved through timing harvest in the spring when growth rates are higher and leaching potential is lower. Based on N uptake rates observed in this study and average pond discharge, a plantation of 5.4 ha would be required for N recovery on a typical dairy farm in New Zealand.
Subject(s)
Cattle , Conservation of Natural Resources , Nitrogen/metabolism , Trees , Agriculture , Animals , Refuse Disposal/methods , Time Factors , Water Movements , Water Pollution/prevention & controlABSTRACT
The effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on the liver of C57BL/6J mice is a model for clinical sporadic porphyria cutanea tarda (PCT). There is massive uroporphyria, inhibition of uroporphyrinogen decarboxylase (UROD) activity, and hepatocellular damage. A variety of evidence implicates the CYP1A2 enzyme as necessary for mouse uroporphyria. Here we report that, 5 weeks after a single oral dose of TCDD (75 microg/kg), Cyp1a2(+/+) wild-type mice showed severe uroporphyria and greater than 90% decreases in UROD activity; in contrast, despite exposure to this potent agent Cyp1a2(-/-) knockout mice displayed absolutely no increases in hepatic porphyrin levels, even after prior iron overload, and no detectable inhibition of UROD activity. Plasma levels of alanine-aminotransferase (ALT) and aspartate aminotransferase (AST)-although elevated in both genotypes after TCDD exposure-were significantly less in Cyp1a2(-/-) than in Cyp1a2(+/+) mice, suggesting that the absence of CYP1A2 also affords partial protection against TCDD-induced liver toxicity. Histological examination confirmed a decrease in hepatocellular damage in TCDD-treated Cyp1a2(-/-) mice; in particular, there was no bile duct damage or proliferation that in the Cyp1a2(+/+) mice might be caused by uroporphyrin. We conclude that CYP1A2 is both necessary and essential for the potent uroporphyrinogenic effects of TCDD in mice, and that CYP1A2 also plays a role in contributing to TCDD-induced hepatocellular injury. This study has implications for both the toxicity assessment of TCDD and the hepatic injury seen in PCT patients.
Subject(s)
Cytochrome P-450 CYP1A2/deficiency , Polychlorinated Dibenzodioxins/toxicity , Porphyrias, Hepatic/enzymology , Porphyrias, Hepatic/prevention & control , Uroporphyrins/urine , Animals , Atrophy/chemically induced , Crosses, Genetic , Cytochrome P-450 CYP1A2/genetics , Cytochrome P-450 CYP1A2/metabolism , Environmental Pollutants/toxicity , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Porphyrias, Hepatic/chemically induced , Thymus Gland/drug effects , Thymus Gland/pathology , Uroporphyrinogen Decarboxylase/metabolism , Uroporphyrins/metabolismABSTRACT
Indomethacin is a prostaglandin synthetase inhibitor sometimes used for tocolysis. Several placebo-controlled trials and trials comparing indomethacin to other potential first-line tocolytic agents support its efficacy for delaying delivery for >48 hours. Recent observational studies, however, have raised concerns about the safety of indomethacin, implicating it with increased rates of intraventricular hemorrhage and necrotizing enterocolitis. Careful analysis of these observational studies suggests that these results should be viewed with caution, because of uncontrolled confounding by indication. A recent decision analysis supports the risk/benefit analysis of indomethacin in this setting. Still, the future of indomethacin in preterm labor should be guided by well-designed prospective clinical trials. Such studies are underway.
Subject(s)
Indomethacin/adverse effects , Indomethacin/therapeutic use , Obstetric Labor, Premature/drug therapy , Tocolysis/methods , Tocolytic Agents/adverse effects , Tocolytic Agents/therapeutic use , Adult , Clinical Trials as Topic , Female , Humans , Indomethacin/pharmacokinetics , Infant, Newborn , Pregnancy , Pregnancy Outcome , Prostaglandins/pharmacology , Prostaglandins/physiology , Risk Assessment , Tocolysis/adverse effects , Tocolytic Agents/pharmacokineticsABSTRACT
The synergistic interaction of iron overload, AHR: genotype and exposure to a mixture of polychlorinated biphenyls (PCBs) (Aroclor 1254) in mice leads to hepatic porphyria, oxidative DNA damage and cancer. In humans, hepatocellular cancer is associated with iron overload and hepatic porphyria. Neither the mechanism of hepatic carcinogenesis induced by PCBs in rodents nor hepatocellular cancer induced by iron and porphyria in humans are understood. To test the hypothesis that chronic interaction of iron and PCBs may induce mutagenesis in liver DNA, lambda /lacI transgenic C57BL/6 mice were given iron dextran (600 mg iron/kg) and then administered Aroclor 1254 in the diet (0.01%) for 7 weeks. Hepatic iron, CYP1A activity and CYP1A1/1A2 protein were elevated >20-fold as a result of iron or Aroclor treatments, respectively, but porphyria with associated histological changes only developed in the combined iron/Aroclor treatment group. lambda/lacI shuttle vectors were isolated from liver genomic DNA and the mutational frequency (MF) in the lacI gene determined. Both iron and Aroclor treatments alone caused significant small increases in MF (1.5- and 1.4-fold, respectively), however, the MF following the combined iron and Aroclor treatment (1. 6-fold) was not greater than the additive effects. In contrast, the MF was significantly elevated (4.7-fold) in liver DNA of mice 2 weeks following five daily doses of N-nitrosodimethylamine (4 mg/kg). These studies demonstrate that neither PCBs nor iron overload caused marked point mutations even in a combination regime that leads to oxidative damage and cancer. There was also no strong evidence either that porphyrins or chronic CYP1A1 expression induced by the PCBs after this period caused marked point mutagens or simple deletions. Hence, to understand the PCBs-iron synergism more complex scenarios than point mutations or simple deletions must be invoked.
Subject(s)
Bacterial Proteins/genetics , Escherichia coli Proteins , Iron Overload/genetics , Liver Neoplasms, Experimental/genetics , Liver/pathology , Mutation , Polychlorinated Biphenyls/toxicity , Porphyrias/genetics , Repressor Proteins/genetics , Animals , Bacteriophage lambda/genetics , Cytochrome P-450 CYP1A1/metabolism , Cytochrome P-450 CYP1A2/metabolism , DNA Damage , Dimethylnitrosamine/toxicity , Genetic Vectors/genetics , Iron Overload/metabolism , Lac Repressors , Liver/drug effects , Liver/metabolism , Liver Neoplasms, Experimental/metabolism , Male , Mice , Mice, Inbred C57BL , Mice, Transgenic , Models, Biological , Mutagenesis , Oxidative Stress/drug effects , Porphyrias/metabolism , Porphyrins/metabolismABSTRACT
Toxic and carcinogenic free radical processes induced by drugs and other chemicals are probably modulated by the participation of available iron. To see whether endogenous iron was genetically variable in normal mice, the common strains C57BL/10ScSn, C57BL/6J, BALB/c, DBA/2, and SWR were examined for major differences in their hepatic non-heme iron contents. Levels in SWR mice were 3- to 5-fold higher than in the two C57BL strains, with intermediate levels in DBA/2 and BALB/c mice. Concentrations in kidney, lung, and especially spleen of SWR mice were also greater than those in C57BL mice. Non-denaturing PAGE of hepatic ferritin from all strains showed a major holoferritin band at approximately 600 kDa, with SWR mice having > 3-fold higher levels than C57BL strains. SDS PAGE showed a band of 22 kDa, mainly representing L-ferritin subunits. A trace of a subunit at 18 kDa was also detected in ferritin from SWR mice. The 18 kDa subunit and a 500 kDa holoferritin from which it originates were observed in all strains after parenteral iron overload, and there was no major variation in ferritin patterns. Although iron uptake studies showed no evidence for differential duodenal absorption between strains to explain the variation in basal iron levels, acquisition of absorbed iron by the liver was significantly higher in SWR mice than C57BL/6J. As with iron and ferritin contents, total iron regulatory protein (IRP-1) binding capacity for mRNA iron responsive element (IRE) and actual IRE/IRP binding in the liver were significantly greater in SWR than C57BL/6J mice. Cytosolic aconitase activity, representing unbound IRP-1, tended to be lower in the former strain. SWR mice were more susceptible than C57BL/10ScSn mice to the toxic action of diquat, which is thought to involve iron catalysis. If extrapolated to humans, the findings could suggest that some people might have the propensity for greater basal hepatic iron stores than others, which might make them more susceptible to iron-catalysed toxicity caused by oxidants.
Subject(s)
Ferritins/metabolism , Iron-Sulfur Proteins/metabolism , Iron/metabolism , Liver/metabolism , Oxidative Stress/physiology , RNA-Binding Proteins/metabolism , Absorption , Animals , Diquat/toxicity , Ferritins/genetics , Genetic Variation , Herbicides/toxicity , Iron Regulatory Protein 1 , Iron-Regulatory Proteins , Iron-Sulfur Proteins/genetics , Mice , Mice, Inbred C57BL , Mice, Inbred DBA , Nonheme Iron Proteins , Oxidative Stress/drug effects , Oxidative Stress/genetics , RNA-Binding Proteins/genetics , Species SpecificityABSTRACT
We show here that DBA/2 strain mice have a complex mutation/polymorphism in the promoter region of the Trp53 locus (the mouse p53 locus). This region has previously been shown to be essential for p53 expression. We further show that the DBA/2 mutation is associated with approximately fourfold lower p53 levels in thymocytes treated with the DNA-damaging agent etoposide in-vitro, and with relative resistance of these thymocytes to apoptosis induced by the DNA-damaging agent etoposide compared with C57BL/6 mice. When part of the promoter containing this mutation was inserted into a plasmid containing a luciferase reporter gene but lacking eukaryote promoter sequences and transfected into MCF-7 human breast cell line cells, the mean luciferase activity was slightly less with the DBA/2 than with the C57BL/6 promoter-reporter construct (p < 0.01). We found that DBA/2xC57BL/6 F2 hybrid mice with the DBA/2 genotype at the Trp53 locus were relatively resistant to tetrachlorodibenzo-p-dioxin toxicity, and this resistance was additive with resistance associated with the Ahr locus. DBA/2 mice are long-lived and do not have particularly high levels of cancer, suggesting either that they carry other compensatory tumour resistance alleles (such as Ahr(d)), or that, while there may be a p53 protein dosage effect for acute toxicity, lower than normal levels of p53 may still be sufficient to protect against cancer. In evolutionary terms, it may be better to maintain low levels of p53 in order to avoid death from acute toxicity, even at the expense of a higher incidence of cancer in later life.
Subject(s)
Genes, p53 , Polychlorinated Dibenzodioxins/toxicity , Animals , Base Sequence , Body Weight/drug effects , DNA , Etoposide/pharmacology , Genotype , Homozygote , Humans , Mice , Mice, Inbred C57BL , Mice, Inbred DBA , Organ Size/drug effects , Polymorphism, Single-Stranded Conformational , Promoter Regions, Genetic , Thymus Gland/cytology , Thymus Gland/drug effects , Tumor Cells, CulturedABSTRACT
The binding of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) with the aryl hydrocarbon (AH) receptor and subsequent changes in gene expression have been studied intensively, but the mechanisms by which these lead to toxicity are unclear. We investigated the influence of iron, previously implicated in TCDD-induced hepatic porphyria, in mice with alleles of Ahr that encode receptors with varied affinity for TCDD. The administration of iron to Ahrb-1 C57BL/6J (AH-responsive) mice before a single dose of TCDD (75 micrograms/kg) markedly potentiated not only the hepatic porphyria but also general hepatocellular damage and elevation of plasma hepatic enzymes. The formation of hydroxylated and peroxylated derivatives of uroporphyrins formed from uroporphyrinogen and the induction of a mu-glutathione transferase (GST) were consistent with the operation of an oxidative mechanism. In a comparison of C57BL/6J mice with Ahrb-2 BALB/c (AH-responsive) and Ahrd SWR and DBA/2 (AH-nonresponsive) mice, iron overcame the weak hepatic porphyria and toxicity responses in BALB/c and SWR strains but not in DBA/2. CYP1A isoforms are strongly implicated in the mechanism of porphyria, but activities were lowered by 20-30% with iron treatment, and a comparison of levels between strains did not fully account for the resistance of DBA/2 mice. Studies with the use of gel shift assays and cytosolic aconitase of the capacity of the iron regulatory protein controlling the translation of some iron metabolism proteins showed a significant difference between C57BL/6J and DBA/2 mice after the administration of TCDD. We conclude that iron potentiates both the hepatic porphyria and toxicity of TCDD in susceptible mice in an oxidative process with disturbance of iron regulatory protein capacity. Iron even overcomes the AH-nonresponsive Ahrd allele in the SWR strain but not in DBA/2 mice, which remain resistant.
Subject(s)
Iron/metabolism , Liver/drug effects , Liver/metabolism , Polychlorinated Dibenzodioxins/metabolism , Polychlorinated Dibenzodioxins/toxicity , Porphyrias, Hepatic/metabolism , Receptors, Aryl Hydrocarbon/genetics , Alleles , Animals , Chromatography, High Pressure Liquid , Cytochrome P-450 CYP1A1/biosynthesis , Drug Synergism , Enzyme Induction , Ferritins/biosynthesis , Glutathione Transferase/biosynthesis , Male , Mice , Mice, Inbred Strains , Oxidation-Reduction , Porphyrinogens/analysis , Porphyrinogens/pharmacokinetics , Protein Biosynthesis , RNA, Messenger/metabolism , Receptors, Transferrin/biosynthesisABSTRACT
Administration in the drinking water of the orally-active iron chelator 1,2-diethyl-3-hydroxypyridin-4-one (CP94) to C57BL/10ScSn mice caused the development of hepatic protoporphyria. This was detected after 1 week and continued as long as the chelator was given (15 weeks). The more hydrophilic 1,2-dimethyl- and 1-hydroxyethyl,2-ethyl-analogues (CP20 and CP102) were also tested, but they were both inactive in inducing accumulation of protoporphyrin in the liver. Restriction of in vivo iron supply for ferrochelatase seemed a likely mode of action, but an approximately 30% decrease in activity of this enzyme was also observed when measured in vitro. Extracts of livers from mice given CP20, CP94, and CP102 showed no potential to inhibit mouse ferrochelatase, in contrast to the findings with an extract from mice treated with the known porphyrogenic chemical 4-ethyl-3, 5-diethoxycarbonyl-2,6-dimethyl-1,4-dihydropyridine, indicating that ferrochelatase inhibition did not occur by the formation of an N-ethyl-protoporphyrin derived from metabolism by cytochrome P450, CP20, CP94, CP102, and CP117 (the pivoyl ester of CP102) all caused significant depression of the levels of ferritin-iron and total nonheme iron, but only CP94 caused the significant accumulation of protoporphyrin. Protoporphyria did not occur with iron overloaded C57BL/10ScSn mice or in SWR mice that had elevated basal iron status. Although the protoporphyrin had only a small effect on the total levels of the hemoprotein cytochrome P450 in C57BL/10ScSn mice, the activity of the CYP2B isoforms of cytochrome P450 was actually induced in both strains. The results show that CP94 could cause protoporphyria in individuals of low iron status, perhaps through specifically targeting particular iron pools available to ferrochelatase and by concomitantly stimulating heme synthesis.
Subject(s)
Iron Chelating Agents/toxicity , Porphyrias, Hepatic/chemically induced , Pyridones/toxicity , Administration, Oral , Animals , Cytochrome P-450 Enzyme System/drug effects , Ferrochelatase/antagonists & inhibitors , Heme/biosynthesis , Iron/metabolism , Iron Chelating Agents/administration & dosage , Liver/drug effects , Liver/enzymology , Liver/metabolism , Male , Mice , Mice, Inbred C57BL , Porphyrias, Hepatic/metabolism , Pyridones/administration & dosageABSTRACT
In mice, depression of hepatic uroporphyrinogen decarboxylase (UROD) leading to porphyrin accumulation (uroporphyria) occurs with chlorinated ligands of the aryl hydrocarbon (AH) receptor especially after iron overload. However, in the absence of chlorinated ligands, iron itself will eventually cause uroporphyria, but this response is not associated with the Ahr genotype. These effects are potentiated by administration of the haem precursor 5-aminolaevulinate (ALA). The aim of this study was to investigate the effects of ALA alone. Prolonged administration of 2 mg ALA/mL in the drinking water to SWR mice also led to decarboxylase insufficiency (11% of control) and uroporphyria by 8 weeks, whereas DBA/2 mice did not show reduced enzyme activity. Both strains are considered AH nonresponsive and analysis of the Ahr gene using restriction fragment length polymorphism was consistent with SWR, like DBA/2, possessing the Ahrd allele. Exposure of isolated hepatocytes to ALA (150-500 microM) for up to 48 hr showed a significant accumulation of both uroporphyrin and coproporphyrin in the medium, which for uroporphyrin particularly was significantly greater with SWR than with DBA/2 cells. Basal in vivo CYP1A2 activity, measured as microsomal methoxyresorufin dealkylation, was significantly greater in SWR than in DBA/2 mice (1.3-fold), but it was unclear whether this was sufficient to explain the marked difference in sensitivities of the two strains. Despite SWR mice being AH nonresponsive, uroporphyria and decarboxylase depression after an initial iron overload and ALA for 3 weeks were greatly potentiated by a single dose (100 mg/kg) of hexachlorobenzene (a weak AH ligand). The results demonstrate that there is a genetic difference in mice independent of the Ahr genotype and response to iron, which influences the susceptibility to ALA-induced uroporphyria. Thus chemicals, iron and ALA can act independently, but also together, to cause porphyria in susceptible individuals.
Subject(s)
Aminolevulinic Acid/toxicity , Porphyrias/chemically induced , Porphyrias/genetics , Receptors, Aryl Hydrocarbon/genetics , Animals , Cytochrome P-450 Enzyme System/metabolism , Genotype , Iron Overload/complications , Liver/drug effects , Liver/enzymology , Male , Mice , Mice, Inbred DBA , Porphyrias/etiology , Species Specificity , Uroporphyrinogen Decarboxylase/metabolismABSTRACT
In Ah-responsive C57BL/10ScSn mice a single dose of iron significantly potentiated the property of the polychlorinated biphenyl (PCB) mixture Aroclor 1254 to induce porphyria by inhibition at the uroporphyrinogen decarboxylase stage of hepatic haem biosynthesis. The induction of liver tumors and other lesions were also enhanced markedly by iron overload suggesting a link between porphyria and cancer. The cellular, molecular and biochemical processes involved have been investigated in attempts to explain these phenomena by an iron-catalysed 'oxidative stress' mechanism.
Subject(s)
Iron/toxicity , Liver Neoplasms, Experimental/chemically induced , Polychlorinated Biphenyls/toxicity , Porphyrias, Hepatic/chemically induced , Animals , Aroclors/toxicity , Drug Synergism , Male , Mice , Mice, Inbred C57BL , Mice, Inbred DBAABSTRACT
The reaction of bovine erythrocyte acetylcholinesterase (acetylcholine acetylhydrolase, EC 3.1.1.7) with a set of structurally related phosphorothiolates was studied in order to investigate the properties of the phosphorylated enzymes and to identify the leaving group. OOS- and OOS-trimethyl phosphorothiolates and their triethyl analogues inhibit acetylcholinesterase reversibly and by progressive inhibition, and the phosphorylated enzymes undergo both spontaneous reactivation and aging. For each compound the enzyme-inhibitor dissociation constant, and the rate constants for inhibition (ka), reactivation and aging have been derived. The OOS-compounds are more potent inhibitors than the OOS-compounds, and the derived inhibited enzymes reactivate and age faster. By comparing reactivation and aging rate constants with those obtained from phosphorylated enzymes of known structure it was concluded that the leaving group of during phosphorylation is the S-alkyl. SSS-trimethyl and -triethyl phosphorothiolates also form reversible complexes and inhibit the enzyme progressively. With these inhibitors the phosphorylated enzymes did not reactivate either spontaneously or in response to oximes under conditions successful for the other inhibitors. The ka values (37 degrees C, pH 7.4) range from 30 M-1 X min-1 (OOS-trimethyl phosphorothiolate) to 6.7 X 10(3) M-1 X min-1 (OOS-triethyl phosphorothiolate) as compared to 1.25 X 10(5) M-1 X min-1 determined for isomalathion (O, S-dimethyl S-(1,2-dicarbethoxyethyl)-phoshporodithioate), which was used as one of the reference compounds. If the inhibitory potency of the trialkyl phosphorothiolates is calculated from measurements made after a fixed preincubation time the results in ka values will be misleading.
Subject(s)
Cholinesterase Inhibitors , Organothiophosphorus Compounds/pharmacology , Acetylcholinesterase/blood , Animals , Cattle , Drug Stability , Enzyme Activation/drug effects , Erythrocytes/enzymology , Kinetics , Malathion/pharmacology , PhosphorylationABSTRACT
1. It was proposed [Johnson (1974) J. Neurochem. 23, 785--789] that both inhibition of neurotoxic esterase of nervous tissue and subsequent 'aging' of the inhibited esterase are necessary events in the pathogenesis of organophosphate-induced delayed neuropathy: aging has now been demonstrated with a number of neurotoxic compounds. 2. Reactivation by KF was observed for hen brain neurotoxic esterase inhibited by 14 organophosphates and phosphonates, and time-dependent loss of reactivatibility (aging) occurred in every case. 3. For five other compounds no reactivation occurred and aging could not therefore be established, but independent evidence for two compounds suggests that aging was rapid. 4. Half-lives of aging of neurotoxic esterase inhibited by phosphates ranged from less than 1 min to 10 min, and for phosphonates the range was 3--600 min. 5. The relationship of these findings to the mechanism of toxicity and to the prospects of therapy are considered. 6. Aging occurred rapidly with aryloxy and linear alkoxy groups attached to phosphorus and slowly with a highly branched alkoxy substituent: these effects seem incompatible with an SN1 (dealkylation) mechanism.
Subject(s)
Brain/enzymology , Esterases/antagonists & inhibitors , Organophosphorus Compounds/pharmacology , Animals , Brain/drug effects , Chickens , Enzyme Activation/drug effects , Fluorides/pharmacology , Half-Life , Neurotoxins/antagonists & inhibitors , Time FactorsABSTRACT
Chymotrypsin and neurotoxic esterase (NTE) have some similarities. After inhibition of concentrated (80-800 micro M) chymotrypsin by aryl saligenin cyclic phosphates it is known that aging occurs and some phenolic material becomes attached to protein. This binding has now been shown to be a manifestation of non-specific reaction with any available electrophile such as Tris, reduced glutathione (GSH), or protein. The reaction is therefore not a model for the 100% efficient transfer of alkyl groups to protein which occurs during aging of NTE inhibited by dialkyl phosphates.
Subject(s)
Benzyl Alcohols/pharmacology , Benzyl Compounds/pharmacology , Central Nervous System/drug effects , Chymotrypsin/analysis , Esterases/analysis , Organophosphorus Compounds/pharmacology , Chymotrypsin/antagonists & inhibitors , Esterases/antagonists & inhibitors , Glutathione/metabolism , Insecticides/toxicity , Models, Chemical , Protein Binding , Time FactorsABSTRACT
1. It was proposed [Johnson (1974) J. Neurochem.23, 785-789] that an essential step in the genesis of delayed neuropathy caused by some organophosphorus esters was aging of phosphorylated neurotoxic esterase, involving generation of a charged monosubstituted phosphoric acid residue on the protein. 2. Neurotoxic esterase of hen brain was inhibited with di-isopropyl phosphorofluoridate either unlabelled or mixed-labelled with (3)H and (32)P. 3. Reactivation of inhibited enzyme by KF was possible only immediately after a brief inhibition:aging at pH8.0 and 37 degrees C occurred with a half-life of about 2-4min. 4. When the radiolabelled enzyme was studied no loss of label was observed during the expected aging period, but a change in the nature of the bound radioisotopes occurred (half-life=3.25min). 5. Alkaline hydrolysis of labelled enzyme liberated di-isopropyl phosphate at early times after labelling, but increasing amounts of monoisopropyl phosphate plus a volatile tritiated compound (possibly propan-2-ol) at later times. 6. Treatment of labelled enzyme with KF released di-isopropyl phosphate and caused reactivation of enzyme to similar degrees. It is concluded that the chemical change from di-isopropyl phosphoryl-enzyme to mono-isopropyl phosphoryl-enzyme and the loss of reactivatibility are related. 7. The rate of aging is similar at pH5.2, 6.5 and 8. Aging is unaffected by addition of reduced glutathione and imidazole at pH5.2 or 8, and none of the transferred (3)H is trapped by these reagents. The mechanism of aging must be different from the better-known dealkylation aging of the cholinesterases.