Studies on the mechanism of toxicity of acetaminophen. Synthesis and reactions of N-acetyl-2,6-dimethyl- and N-acetyl-3,5-dimethyl-p-benzoquinone imines.

N-Acetyl-2,6-dimethyl-p-benzoquinone imine and N-acetyl-3,5-dimethyl-p-benzoquinone imine were prepared from 2,6-dimethylacetaminophen and 3,5-dimethylacetaminophen by oxidation with lead tetraacetate. Reaction of N-acetyl-2,6-dimethyl-p-benzoquinone imine with hydrochloric acid gave 3'-chloro-2',6'-dimethyl-4'-hydroxyacetanilide, whereas ethanethiol, aniline, and ethanol gave tetrahedral adducts resulting from addition to the imine carbon. Water gave 2,6-dimethyl-p-benzoquinone. With N-acetyl-3,5-dimethyl-p-benzoquinone imine, water and aniline gave substitution on the imine carbon, yielding 2,6-dimethyl-p-benzoquinone and 3,5-dimethyl-N-phenyl-p-benzoquinone imine, respectively. Ethanethiol gave 3'.5'-dimethyl-2'-(ethylthio)-4'-hydroxyacetanilide. The toxicity of 2,6-dimethylacetaminophen and 3,5-dimethylacetaminophen was examined histologically in mice and rats. 3,5-Dimethylacetaminophen was slightly more nephrotoxic but showed a similar hepatotoxicity to acetaminophen. 2,6-Dimethylacetaminophen, like N-methylacetaminophen, showed very little tissue damage.

N-hydroxyacetaminophen: a postulated toxic metabolite of acetaminophen.

The decomposition of N-hydroxyacetaminophen has been shown to occur via an initial first-order dehydration step to N-acetyl-p-benzoquinone imine with a rate constant at pH 7.6 of 8.66 x 10(-3) min-1 and a half-life of 80 min. This is followed by a complex reaction between the quinone imine and the N-hydroxy compound to ultimately yield p-nitrosophenol and acetaminophen. The glucuronide and sulfate conjugates of N-hydroxyacetaminophen have been observed as urinary metabolites of N-hydroxyacetaminophen. No N-hydroxylated metabolites were found among the metabolites of acetaminophen. These results have been interpreted to show that N-hydroxyacetaminophen is not a metabolite of acetaminophen. It is proposed that the hepatotoxicity and nephrotoxicity of acetaminophen are mediated by a direct oxidation of acetaminophen to the toxic reactive intermediate N-acetyl-p-benzoquinone imine by the cytochrome P450 dependent mixed-function oxidase system.

Primary brain trauma in non-accidental injury.

The brains from 12 babies up to 21/2 years of age, who died after repeated non-accidental injury to the head, were subjected to detailed neuropathological examination. The nine brains from infants under 5 months showed contusional tears--slit like lesions in the white matter surrounded by astrocytes and associated with evidence of old and recent haemorrhage. The three brains from infants over 5 months showed white matter lesions similar to those seen in adults after closed head injury, including damage in the dorsolateral quadrant of the brain stem without axonal hemispheric damage, which may have been a result of whiplash injury after shaking. In addition, all the brains examined showed diffuse gliosis. This paper draws attention to contusional tears and other white matter lesions, which the authors believe are manifestations of mechanical damage produced by trauma. The long term neurological and intellectual defects observed in patients suffering non-accidental injury early in life are increasingly being recognised, although it is difficult to identify the extent to which these are due to social or neuropathological factors. We suggest that the white matter damage we describe has an important role.

Clofibrate-induced in vitro hepatoprotection against acetaminophen is not due to altered glutathione homeostasis.

Prior induction of peroxisome proliferation protects mice against the in vivo hepatotoxicity of acetaminophen and various other bioactivation-dependent toxicants. The mechanisms underlying such chemoresistance are poorly understood, although they have been suggested to involve alterations in glutathione homeostasis. To clarify the role of glutathione in this phenomenon, we isolated hepatocytes from mice in which hepatic peroxisome proliferation had been induced with clofibrate. The cells were incubated with a range of acetaminophen concentrations and the extent of cell killing after up to 8 h was assessed by measuring lactate dehydrogenase leakage from the cells. Hepatocytes from clofibrate-pretreated mice were much less susceptible to acetaminophen than cells from vehicle-treated controls. However, the extent of glutathione depletion during exposure to acetaminophen was similar in both cell types, as were rates of excretion of the product of glutathione-mediated detoxication of acetaminophen's quinoneimine metabolite, 3-glutathionyl-acetaminophen. The glutathione-replenishing ability of clofibrate-pretreated cells after a brief exposure to diethyl maleate also resembled that of control cells. More importantly, prior depletion of glutathione by diethyl maleate did not abolish the resistance of clofibrate-pretreated cells to acetaminophen. Taken together, these findings indicate that although glutathione-dependent pathways may contribute to hepatoprotection during peroxisome proliferation, the resistance phenomenon is not due exclusively to this mechanism.

Tropospheric ozone: respiratory effects and Australian air quality goals.

OBJECTIVE: To review the health effects of tropospheric ozone and discuss the implications for public health policy. DESIGN: Literature review and consultation with scientists in Australia and overseas. Papers in English or with English language abstracts were identified by Medline search from the international peer reviewed published reports. Those from the period 1980-93 were read systematically but selected earlier papers were also considered. Reports on ozone exposures were obtained from environmental agencies in the region. RESULTS: Exposure to ozone at concentrations below the current Australian air quality goal (0.12 ppm averaged over one hour) may cause impaired respiratory function. Inflammatory changes in the small airways and respiratory symptoms result from moderate to heavy exercise in the presence of ozone at levels of 0.08-0.12 ppm. The changes in respiratory function due to ozone are short lived, vary with the duration of exposure, may be modified by levels of other pollutants (such as sulphur dioxide and particulates), and differ appreciably between individuals. Bronchial lavage studies indicate that inflammation and other pathological changes may occur in the airways before reductions in air flow are detectable, and persist after respiratory function has returned to normal. It is not known whether exposures to ozone at low levels (0.08-0.12 ppm) cause lasting damage to the lung or, if such damage does occur, whether it is functionally significant. At present, it is not possible to identify confidently population subgroups with heightened susceptibility to ozone. People with asthma may be more susceptible to the effects of ozone than the general population but the evidence is not consistent. Recent reports suggest that ozone increases airway reactivity on subsequent challenge with allergens and other irritants. Animal studies are consistent with the findings in human populations. CONCLUSION: A new one hour air quality ozone goal of 0.08 ppm for Australia, and the introduction of a four hour goal of 0.06 ppm are recommended on health grounds.

Hyaluronidase and sodium hyaluronate in cataract surgery.

The use of sodium hyaluronate in cataract surgery and intraocular lens implantation is often followed by a postoperative rise of intraocular pressure. A trial is described in which 10 patients underwent bilateral cataract extraction and Binkhorst intraocular lens implantation with the use of sodium hyaluronate. The enzyme hyaluronidase was instilled into the anterior chamber of the right eye only, to aid removal of sodium hyaluronate, and resulted in a statistically significant lowering of postoperative intraocular pressure in right eyes compared with left. Other uses of the enzyme are discussed.

The influence of metabolic variation on analgesic nephrotoxicity. Experiments with the Gunn rat.

The analgesics aspirin and paracetamol administered as single I.V. doses produce renal lesions in the homozygous Gunn rat. The lesions affect both cortex and medulla but are less severe than the renal lesions of analgesic nephropathy. By contrast the reactive compounds p-aminophenol and 5-aminosalicylic acid which are known to cause renal damage in other less susceptible strains respectively produce cortical and medullary renal lesions in homozygous Gunn rats which are as extensive as those found in patients with analgesic nephropathy. The increased frequency of renal lesions from the analgesics aspirin and paracetamol as compared to heterozygous and albino rats and the increased severity of the lesions due to p-aminophenol and 5-aminosalicylic acid is considered to be at least partly due to impaired glucuronide formation and consequent delayed excretion of nephrotoxic substances.

Hepatotoxicity of phenacetin and paracetamol in the Gunn rat.

Both phenacetin and paracetamol produce acute centrilobular liver necrosis in the homozygous Gunn rat. Paracetamol is more hepatotoxic than phenacetin, and both are more hepatotoxic to the homozygous Gunn rat than to the heterozygous Gunn rat or to the albino rat. These findings have relevance to the role of the compounds in the clinical syndromes of paracetamol induced liver necrosis and analgesic nephropathy.

Neoplasia in the rat induced by N-hydroxyphenacetin, a metabolite of phenacetin.

N-hydroxyphenacetin, a phenacetin metabolite, was fed to rats as a 0.05-0.5% dietary supplement. After 9 months, tumours of the liver were found in 36 of 64 animals. One animal also developed a renal tumour. No tumours were found in control animals. The findings implicate phenacetin as a carcinogen and suggest that N-hydroxyphenacetin may be the metabolite responsible.

The nephrotoxicity of p-aminophenol. I. The effect on microsomal cytochromes, glutathione and covalent binding in kidney and liver.

p-Aminophenol administration lowered the microsomal cytochrome P-450 and b5 content and decreased the activity of NADPH cytochrome c reductase in kidney, but not in liver. Kidney GSH was depleted to 29% of the control value at 2 h, and only partly restored (50% of control) at 24 h. Liver GSH was transiently decreased, the lowest levels (77% of control) occurring at 30 min. The maximum level of covalently bound radioactivity was at two hours when 16.8% of the total radioactivity in kidney, 1.5% in liver and 3.6% in plasma was protein bound. At this time 81% of the total radioactivity in kidney and 95% of that in the liver was present in the soluble fraction.

The nephrotoxicity of p-aminophenol. II. The effect of metabolic inhibitors and inducers.

Inducers and inhibitors of the microsomal mixed function oxidase system have no consistent effect upon the nephrotoxicity of p-aminophenol, or on binding of the compound in vivo to cell protein. p-[ring-3H]Aminophenol was bound in vitro to kidney microsomal protein and to a lesser extent to liver. The binding was enhanced by preincubation of the p-aminophenol in air and inhibited by ascorbate, GSH, N2 and NADPH. These findings indicate that in contrast to paracetamol hepatoxicity which is dependent upon the mixed function oxidase system, that nephrotoxicity of p-aminophenol is dependent upon oxidation to a toxic metabolite by some other pathway. A similar metabolite may be responsible for the nephrotoxic action of phenacetin.

Nystagmus and joint position sensation: their importance in posterior occipitocervical fusion in rheumatoid arthritis.

It is widely believed that brain stem dysfunction and cranial nerve palsies in patients with rheumatoid arthritis (RA) are common and related to the vertical translocation of the odontoid process. In our database of 235 patients with seropositive RA and craniocervical junction involvement, we have found a very low incidence of such problems. Long tract signs were common, but loss of proprioception (joint position sensation) as the sole neurologic deficit was rare. Nystagmus was found to be associated with the tonsillar herniation of a Chiari 1 malformation and loss of joint position sensation with severe compression of the posterior aspect of the spinal cord at the craniocervical junction. The implications for posterior occipitocervical fusion, particularly by sublaminar wiring, are discussed.

Hepatic peroxisome proliferation in rodents and its significance for humans.

Peroxisomes are subcellular organelles found in all eukaryotic cells. In the liver they are usually round and measure about 0.5-1.0 microns; in rodents they contain a prominent crystalloid core, but this may be absent in newly formed rodent peroxisomes as well as in human peroxisomes. A major role of the peroxisomes is the breakdown of long-chain fatty acids, thereby complementing mitochondrial fatty-acid metabolism. Many chemicals are known to increase the number of peroxisomes in rat and mouse hepatocytes. This peroxisome proliferation is accompanied by replicative DNA synthesis and liver growth. No clear structure-activity relationships are apparent. Many of these peroxisome proliferators contain acid functions that can modulate fatty acid metabolism. Two mechanisms have been proposed for the induction of peroxisome proliferation. One is based on the existence of one or several specific cytosolic receptors that bind the peroxisome proliferator, facilitating its translocation to the cell nucleus and the activation of the expression of specific genes. The second, perhaps more general, hypothesis involves chemically mediated perturbation of lipid metabolism. These two hypotheses are not mutually exclusive. Many peroxisome proliferators have been shown to induce hepatocellular tumours, despite being uniformly non-genotoxic, when administered at high dose levels to rats and mice for long periods. Three mechanisms have been proposed to explain the induction of tumours. One is based on increased production of active oxygen species due to imbalanced production of peroxisomal enzymes; it has been proposed that these reactive oxygen species cause indirect DNA damage with subsequent tumour formation. In rodents, an alternative mechanism is the promotion of endogenous lesions by sustained DNA synthesis and hyperplasia. Thirdly, it is conceivable that sustained growth stimulation may be sufficient for tumour formation. Marked species differences are apparent in response to peroxisome proliferations. Rats and mice are extremely sensitive, and hamsters show an intermediate response while guinea pigs, monkeys and humans appear to be relatively insensitive or non-responsive at dose levels that produce a marked response in rodents. These species differences may be reproduced in vitro using primary culture hepatocytes isolated from a variety of species including humans. The available experimental evidence suggests a strong association and a probable casual link between peroxisome-proliferator-elicited liver growth and the subsequent development of liver tumours in rats and mice. Since humans are insensitive or unresponsive, at therapeutic dose levels, to peroxisome-proliferator-induced hepatic effects, it is reasonable to conclude that the encountered levels of exposure to these non-genotoxic agents do not present a hepatocarcinogenic hazard to humans.(ABSTRACT TRUNCATED AT 400 WORDS)

One-stage transoral decompression and posterior fixation in rheumatoid atlanto-axial subluxation.

An operation which combined anterior transoral decompression with posterior occipitocervical fixation was used in 68 rheumatoid patients with irreducible anterior neuraxial compression at the craniocervical junction. Fibre-optic laryngoscopy with nasotracheal intubation was less hazardous than tracheostomy. The patients underwent surgery in the lateral position to allow access both to the mouth and to the back of the neck without moving the head. Specially designed instruments allowed visualisation from the front without dividing the soft palate. Posterior stabilisation was achieved by a preformed contoured loop fixed to the occiput, the atlas and the axis by sublaminar wires. The procedure allowed immediate mobilisation and had a very low morbidity in such ill patients.

Dysbarism. A review.

This review of dysbarism outlines the development of the knowledge of the effects of pressure changes on tissues and organs, which is related to a complex of physical, physiological and pharmacological changes. It also shows that with the ever increasing pressures to which man is subject the effects can be regarded as total body rather than the traditional concept of a few target organs.

A method for investigating specialised accidents with special reference to diving.

This paper describes a format for the investigation of complicated accidents which result in diving deaths. It emphasises the necessity for communication between technical, medical and legal personnel to arrive at a reasoned appreciation of factors leading to an accident. Properly applied this can result in a meaningful accumulation of data, which can be periodically analysed and be used in formulating Regulations for prevention of further accidents.

Experiments using high pressure fluid jets on human tissues.

The object of the experiments was to observe the effects of small jets of high pressure fluid on skin and subcutaneous tissues. This was to simulate a situation which might occur following minor failures in the pressure hulls of submersibles. It was found that the dissipation of Kinetic Energy on a small experimental scale resulted in considerable tissue damage. Preliminary tests interspersing fabric between the jet and skin did not show a great reduction in potential damage. An important aspect considered is the possible introduction of pathogens into tissues when sea water is the fluid.

Dorsal column stimulation (DCS) in chronic pain: report of 31 cases.

Thirty-one patients of chronic pain treated with dorsal column stimulation (DCS) are reported. All of them had been treated previously with drugs and multiple procedures including injections and frequently several operations. After a trial of percutaneous DCS, permanent implantations were carried out. The patients have been followed for up to eight years. Overall, sixty per cent of patients had good to fair relief of pain with DCS. Some of them had a good response for five years and more.

Medical management of active chronic otitis media: a controlled study.

About 2 per cent of adults have active chronic otitis media, the majority being managed by medical means. Previous controlled studies have been unable to show benefit from any medication, including systemic or topical antibiotics, but the effect of the addition of topical steroids to the latter has never been evaluated. One hundred and sixty three adults with active chronic otitis media were randomly allocated to receive either antibiotic/steroid ear drops or placebo therapy over a 4-6 week period. Fifty-two per cent of ears receiving active therapy, as opposed to 30 per cent on placebo therapy (p less than 0.05), became otoscopically inactive if compliance to medication was greater than 70 per cent. However, when there was an open mastoid cavity, active therapy was no more successful than placebo. Though gentamicin was the antibiotic used, there was no evidence of ototoxic inner ear damage. Surprisingly, correlation between clinical activity and patient report of a discharge was poor. Forty per cent of both treatment groups considered that their ear had become dry following therapy and these were not the same patients whose ears had become otoscopically inactive.