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Early prediction of the outcome of a paracetamol overdose based on an analysis of 163 patients.

Clinical and biochemical data obtained from 163 patients who had taken an overdose of paracetamol were examined to determine which factors or measurements were of value in predicting the severity of ensuing liver damage early after ingestion of tablets. Although the overall severity of hepatic necrosis was found to increase with the dose of paracetamol ingested, correlation was not sufficiently close to provide an accurate prognostic index in individuals. Severe hepatic damage was less likely if the patient had vomited or had a stomach wash-out within 6 hr of overdose. The plasma concentrations of paracetamol, measured at known times after overdose, distinguished those who developed hepatic dysfunction from those who did not, but there was a poor correlation, particularly in the first 6 hr after ingestion of tablets, between these values and the severity of ensuing liver damage. Estimates of early plasma paracetamol half-lives from three or more samples taken within 4 hr of admission showed that all patients developing moderate or severe liver damage had half-lives greater than 4 hr, but this was also the case in nearly one-third of those with minimal liver lesions only. It is concluded that there is no completely reliable early prognostic test for individual patients with paracetamol overdose. If each patient is selected for treatment with cysteamine (mercaptamine) or other agents on the basis of plasma paracetamol levels, up to 30% may receive this agent who are at risk from trivial hepatic damage only.

Acetaminophen↗

Reversal of fungitoxicity of 8-quinolinols and their copper (II) bischelates. I. Amino acids and related compounds.

The effect of amino acids and related compounds on the toxicity of 8-quinolinols and their copper (II) bischelates to Aspergillus oryzae (ATCC 1011) Was studied. None of the compounds tested except the thiol-containing compounds, cysteine, cysteamine, glutathione, and N-acetylcysteine reversed the inhibitory action of 8-quinolinol but not that of 5-iodo-8-quinolinol or bis (8-quinolinolato) copper (II). It appears that the mechanism(s) of fungitoxicity of 8-quinolinol is different from that of 5-iodo-8-quinolinol or bis(8-quinolinolato) copper (II.).

Acetylcysteine↗

Threonine metabolism in isolated rat hepatocytes.

The removal of the 1-carbon of threonine can occur via threonine dehydrogenase or threonine aldolase, this carbon ending up in glycine to be liberated by the mitochondrial glycine cleavage system and producing CO(2). Alternatively, in the threonine dehydratase pathway, the 1-carbon ends up in alpha-ketobutyrate, which is oxidized in the mitochondria to CO(2). Rat hepatocytes, incubated in Krebs-Henseleit medium, were incubated with 0.5 mM L-[1-(14)C]threonine, and (14)CO(2) production was measured. Added glycine (0.3 mM) marginally suppressed threonine oxidation. Cysteamine (0.5 mM), a potent inhibitor of the glycine cleavage system, reduced threonine oxidation to 65% of controls. However, alpha-cyanocinnamate (0.5 mM), a competitive inhibitor of mitochondrial alpha-keto acid uptake, reduced threonine oxidation to 35% of controls. These data provided strong evidence that approximately 65% of threonine oxidation occurs through the glycine-independent threonine dehydratase pathway. Glucagon (10(-7) M) increased threonine oxidation and stimulated threonine uptake by these cells. In summary, the majority of threonine oxidation occurs through the threonine dehydratase pathway in rat hepatocytes, and threonine oxidation is increased by glucagon, which also increases threonine's transport.

Animals↗

Somatostatin mediates bombesin inhibition of chloride secretion by rectal gland.

The function of bombesin-like peptide, a neurotransmitter present in nerve fibers of elasmobranch rectal glands, is unknown. Since the principal activity of the rectal gland is to secrete chloride, the effects of bombesin on chloride secretion and the role of somatostatin in this response was studied. Bombesin failed to stimulate secretion in rectal glands perfused in the basal state. When added to glands stimulated by a constant infusion of vasoactive intestinal peptide (VIP), bombesin (8 x 10(-7) M) reversibly inhibited chloride secretion by 56 +/- 9.7% and at the same time evoked a 10-fold increase in the liberation of somatostatin into the venous effluent. Inactivation of somatostatin by the addition of cysteamine partially suppressed the inhibitory effect of bombesin on glandular secretion. The effect of bombesin to reduce chloride secretion was completely prevented by the calcium channel blocker nifedipine, which inhibits neurotransmitter release. These results suggest that bombesin inhibits the effect of VIP to stimulate chloride secretion by releasing somatostatin from neurosecretory nerve terminals within the rectal gland.

Animals↗

Evidence for a central inhibitory role of growth hormone secretagogues and ghrelin on gastric acid secretion in conscious rats.

We examined the possible central and peripheral effects of synthetic growth hormone secretagogues (GHS), hexarelin (Hexa) and EP 40737 (D-Thr-D-Trp (2-Me)-Ala- Trp-D-Phe-Lys-NH2), and of their endogenous counterpart, ghrelin, on gastric acid secretion. The compounds were administered intracerebroventricularly (i.c.v.) or subcutaneously (s.c.) in conscious male rats and the volume of gastric secretion and gastric acid output were examined 3 h after pylorus ligation (Shay-test). Central Hexa, EP 40737 and ghrelin administration (from 0.1 pmol to 1 nmol/rat, i.c.v.) significantly inhibited gastric acid secretion. The maximum inhibitory effect on gastric acid output was detected at the dose of 10 pmol/rat, i.c.v. for Hexa (-51.3%), of 100 pmol/rat, i.c.v. for EP 40737 (-70%) and of 1 pmol/rat, i.c.v. for ghrelin (-60%). All peptides were less effective at the highest dose used (1 nmol/rat, i.c.v.). Hexa, EP 40737 and ghrelin injected s.c. did not modify gastric acid secretion. The inhibitory action of Hexa on gastric acid secretion seems to involve brain somatostatinergic system since Hexa (10 pmol/rat, i.c.v.) did not inhibit gastric acid secretion in rats pretreated (4 h before) with cysteamine (300 mg/kg, s.c.), a depletor of endogenous somatostatin. These results show that synthetic GHS and ghrelin exert a central long-lasting inhibitory effect on gastric acid secretion in conscious pylorus-ligated rats. The fact that very low doses of ghrelin and GHS inhibit gastric secretion, provide evidence for a tonic inhibitory role of the peptides in the central control of gastric secretory function.

Animals↗

Role of porphyrin sequestration in the biogenesis of iron-laden astrocytic inclusions in primary culture.

Astrocytes in subcortical regions of the mammalian brain progressively accumulate iron-rich, autofluorecent cytoplasmic inclusions as a function of aging. Cysteamine (CSH) accelerates the appearance of this senescent glial phenotype in situ and in primary rat astroglial cultures. Porphyrins have been implicated as the source of orange-red autofluorescence in these glial inclusions. Yet, CSH has been shown to suppress porphyrin-heme biosynthesis in cultured astroglia. To determine whether porphyrin biosynthesis or sequestration participates in the biogenesis of these glial inclusions, the porphyrin precursor, (3)H-delta-aminolevulinic acid ((3)H-ALA) was administered to CSH-exposed and control rat astroglial cultures followed by light and electron microscopic autoradiography. Control cultures exhibited faint orange-red autofluorescence, intense (3)H-ALA labeling, numerous normal mitochondria and few cytoplasmic inclusions. In these cells, (3)H-ALA labeling largely occurred over normal mitochondria. The CSH-treated astroglia exhibited diminished (3)H-ALA labeling and contained numerous orange-red autofluorescent inclusions. The latter manifested internal compartments delimited by double membranes characteristic of damaged mitochondria. The complement of normal mitochondria in the CSH-exposed cells was markedly reduced. In the CSH-treated cells, (3)H-ALA labeling predominated over the large multi-compartmental inclusions. CSH attenuates de novo porphyrin-heme biosynthesis in astroglia but may induce punctate orange-red autofluorescence in the cytoplasm of these cells by promoting large numbers of damaged, porphyrin-containing mitochondria to form tight aggregates within the nascent gliosomes.

Aminolevulinic Acid↗

Somatostatin in the hippocampus mediates dexamethasone-induced suppression of corticosterone secretion in the rat.

We evaluated the effect of different manipulations of central somatostatin (SS) neurons on dexamethasone (Dex) induced suppression of the hypophyseal-pituitary axis. The Dex suppression test was performed in male rats by evaluating the reduction of plasma levels of corticosterone occurring 4 h after administration of Dex. Administration of cysteamine, a depletor of SS stores, or passive immunization by intracerebroventricular administration of a saturating dose of anti-SS gamma-globulins completely suppressed the ability of Dex to reduce plasma corticosterone levels. Additionally, anti-SS gamma-globulins, infused under stereotaxic control into two discrete areas of the hippocampus (CA3 and DG), completely blocked Dex-induced inhibition of corticosterone secretion. These data indicate that hippocampal SS neurons play a role in the negative feedback regulation of glucocorticoid secretion.

Animals↗

Enhanced tuberoinfundibular dopaminergic neuron activity in thyroidectomized, ovariectomized and estrogen-treated rats with hyperprolactinemia.

The involvement of the tuberoinfundibular dopaminergic (TIDA) neurons in the genesis of hyperprolactinemia in female thyroidectomized rats treated with estrogen was the focus of this study. Rats that were ovariectomized (3 weeks), thyroidectomized (2 weeks) and treated with estrogen for 6 days had a 5- to 10-fold increase in serum prolactin (PRL) levels, while the rats receiving the same treatments but without estrogen had lower PRL levels. The activity of TIDA neurons, using dihydroxyphenylacetic acid (DOPAC) concentration or the DOPAC/dopamine (DA) ratio in the median eminence (ME) as an index, was increased and decreased in estrogen-treated and untreated rats, respectively. The increases in serum PRL level and the activity of TIDA neurons were dependent on the duration of thyroidectomy and could both be lowered by daily injection of thyroid hormone (20 or 100 micrograms/kg b.w. thyroxine, i.p.) for 12 days in a dose-dependent manner. Using dihydroxyphenylalanine (DOPA) accumulation in the ME as another index, we also found an increased DOPA/DA ratio in estrogen-treated hypothyroid rats, which effect could be reversed by replacement of thyroid hormone. Furthermore, treatments with bromocryptine (3 mg/kg b.w./day, s.c.) for 3 days or with cysteamine (100 mg/kg b.w., s.c.) at 26, 15 and 2 h before the rats were sacrificed lowered both serum PRL level and TIDA neuron activity. Since PRL can exert a short-loop feedback control on its own secretion, these results indicate that the increased serum PRL levels in estrogen-treated hypothyroid female rats should be the cause, but not the result, of the increased activity of TIDA neurons.

3,4-Dihydroxyphenylacetic Acid↗

Role of glutathione status in protection against ethanol-induced gastric lesions.

The role of glutathione status in gastric mucosal cytoprotection has been a subject of controversy. Cysteamine, an exogenous sulfhydryl agent and diethyl maleate (DEM), an endogenous glutathione (GSH) depletor both appear to protect rats from ethanol-induced gastric lesions. In this study, we used various agents to alter gastric mucosal GSH levels and assessed the effects on susceptibility to ethanol injury. We found that DEM and buthionine sulfoximine both depleted gastric GSH but only DEM protected against ethanol-induced gastric lesions. L-Oxothiazolidine-4-carboxylate (OXT) and N-acetyl-L-cysteine (NAC) both potentiated ethanol-induced gastric lesions even though only NAC significantly raised the GSH level. The depletion of GSH by DEM was reversed by supplying cysteine in the form of OXT or NAC so that the net result was a GSH level close to normal control. The potentiation of ethanol injury by NAC and OXT was still apparent. These experiments show no relation between gastric GSH levels and susceptibility to ethanol injury.

Acetylcysteine↗

The effect of the new H2-receptor antagonist mifentidine on gastric secretion, gastric emptying and experimental gastric and duodenal ulcers in the rat: comparison with cimetidine and ranitidine.

The new H2-receptor antagonist mifentidine (DA 4577) was tested for its antisecretory and gastric motor effects in comparison with cimetidine and ranitidine. The Shay rat preparation (5 h) was used for studying gastric secretion; the gastric emptying of a liquid meal was chosen for studying gastric motility. All the three compounds inhibited acid secretion in a dose-dependent fashion. Calculated ED50s were 2.3, 12.2 and 92.8 mg X kg-1 for mifentidine, ranitidine and cimetidine, respectively. Therefore, in this animal model, mifentidine was about 40 times more potent than cimetidine and 5 times more potent than ranitidine. As far as gastric emptying is concerned, the effect of equiactive antisecretory doses (i.e. the respective ED50s calculated from the previously established dose-response curves) of all the three antagonists was completely different. Cimetidine delayed emptying rate, whereas ranitidine accelerated it and mifentidine was completely ineffective. However, at higher doses, also this compound affected emptying rate by reducing it dose-dependently. Gastric and duodenal ulcers were induced in the rat by dimaprit (100 mg X kg-1 intravenously) and cysteamine (250 mg X kg-1 subcutaneously), respectively. As far as gastric ulcer is concerned, the ED50s (the effective dose which protected 50% of the animals from lesions) were 0.23, 4.40 and 9.70 mg X kg-1 for mifentidine, ranitidine and cimetidine, respectively. As regards duodenal ulcer, the ED50 was 4.48 for mifentidine and 150.00 mg X kg-1 for ranitidine. In this animal model, the efficacy of cimetidine was very low. Therefore an ED50 could not be determined. In conclusion, results of the present investigation demonstrated that mifentidine is a potent antisecretory compound and an effective anti-ulcer agent in the rat.

Animals↗

Effect of duodenal ulcerogens mepirizole and propionitrile on small intestinal and liver alkaline phosphatase activity in rats.

Duodenal ulcer could be induced in rats by a single subcutaneous injection of the anti-inflammatory agent mepirizole (M) or by the alkyl chemical propionitrile (P). Contrary to M, P provokes HCl hypersecretion as well. We used these animal models of duodenal ulcer to study the preulcerogenic molecular changes in the mucosal cells and their causal relationship to HCl hypersecretion. It was found that M and P induced the dose- and time-dependent decrease of duodenal alkaline phosphatase (DAP) activity. The decrease was detected at 4 h, with a nadir at 12 h after the injection of both drugs. The decrease of alkaline phosphatase activity was organ-specific after P administration and it was even regional-specific along the small intestine after M administration. At the level of mucosal cells of duodenum the effect of the ulcerogens was enzyme-selective. Both ulcerogens decreased protein and alkaline phosphatase activities; however, they had no effect on lysosomal acid phosphatase. Contrary to cysteamine (C), the effect of M and P on DAP depletion could not be reproduced under in vitro conditions. An interference of P and a slight additive effect of C on DAP depletion after simultaneous subcutaneous administration with M to the rats was found. The results indicate that the DAP depletion after in vivo administration of the three duodenal ulcerogens could be provoked by at least two different mechanisms. The decrease of DAP activity seems to be a general property of the duodenal ulcerogens independent of their effects on gastric acid secretion or on the suppression of alkaline secretion in the duodenum. As a late molecular event most probably elicited by the early morphological changes, the decrease of DAP activity could rather be related to ulcer healing than to its pathogenesis.

Alkaline Phosphatase↗

Enzymatic oxidation of mercaptoethanol to isethinic acid and isethionic acid.

The enzymatic oxidation of mercaptoethanol by purified cysteamine oxygenase has been studied. Products were identified by chromatography as isethinic acid (2-hydroxyethan-sulfinic acid) and isethionic acid. Other features of the reaction, as cofactor requirement, the influence of the enzyme concentration on the stage of oxidation, the biological significance of this reaction are also discussed.

Alkanesulfonates↗

Studies of microsomal glucose-6-phosphatase on liver of irradiated rats.

The activity of microsomal glucose-6-phosphatase (EC 3.1.3.9) on male rat liver was measured 1-9 days after whole-body gamma-irradiation. A marked fall of activity, expressed per whole liver, was observed reaching a minimum on the 4th day following irradiation. The enzyme activity is partially and momentarily restored (on day 7), before a new decrease occurred. Furthermore, when the results are expressed per milligram of microsomal proteins, there was no change. Cysteamine, when injected in vivo, kept up the glucose-6-phosphatase of whole liver. On day 4, a histochemical demonstration of the enzyme in liver cells is in accordance with enzyme measures. These observations suggested that the enzyme quantity was altered during the acute radiation syndrome in the rat.

Animals↗

A novel electrophoretic variant of human apolipoprotein E. Identification and characterization of apolipoprotein E1.

A new apolipoprotein E (apo E) phenotype has been demonstrated in a Finnish hypertriglyceridemic subject (R.M.). At the time of this study, R.M.'s plasma triglyceride and cholesterol levels were 1,021 and 230 mg/dl, respectively. The subject's apo E isoelectric focusing pattern was characterized by two major bands, one in the E3 position and the other in the E1 position. Normally the E1 position is occupied by sialylated derivatives of apo E4, E3, or E2. The E1 band of subject R.M. is not a sialylated form, however, because it was not affected by neuraminidase digestion. The identity of the E1 variant as a genetically determined structure was established by amino acid and partial sequence analyses, confirming that the variant is an example of a previously uncharacterized apo E phenotype, E3/1. Both cysteamine modification and amino acid analysis demonstrated that this variant contains two cysteine residues per mole. Sequence analysis of two cyanogen bromide fragments and one tryptic fragment of the apo E3/1 showed that it differs from E2(Arg158----Cys) at residue 127, where an aspartic acid residue is substituted for glycine. This single amino acid interchange is sufficient to account for the one-charge difference observed on isoelectric focusing gels between E2(Arg158----Cys) and the E1 variant. The variant has been designated E1 (Gly127----Asp, Arg158----Cys). When compared with apo E3, the E1 variant demonstrated reduced ability to compete with 125I-LDL for binding to LDL (apo B,E) receptors on cultured fibroblasts (approximately 4% of the amount of binding of apo E3). This defective binding is similar to that of E2-(Arg158----Cys). Therefore, the binding defect of the variant is probably due to the presence of cysteine at residue 158, rather than aspartic acid at residue 127. In contrast, the apo E3 isoform from this subject demonstrated normal binding activity, indicating that it has a normal structure. In family studies, the vertical transmission of the apo E1 variant has been established. It is not yet clear, however, if the hypertriglyceridemia observed in the proband is associated with the presence of the E1(Gly127----Asp, Arg158----Cys) variant.

Adolescent↗

Antioxidants suppress mortality in the female NZB x NZW F1 mouse model of systemic lupus erythematosus (SLE).

Inflammation produces reactive oxygen intermediates (ROI) that cause vascular damage and activate T lymphocytes. Conversely, antioxidants not only protect tissue from oxidative damage but also suppress immune reactivity. The objective of this study was to examine immunomodulatory effects of the non-enzymatic antioxidants, N-acetylcysteine (NAC) and cysteamine (CYST), on autoimmune disease, glomerulonephritis, and mortality in the female B/W mouse model of human systemic lupus erythematosus (SLE). The development of murine lupus was assessed during the lifespan of female B/W mice given NAC or CYST. Morbidity and mortality were assessed daily. At 6 week intervals mice were examined for weight change, albuminuria, serum BUN, antibodies to DNA, and IgG immunoglobulin levels. Serum prolactin, estrogen and progesterone were measured at 18 weeks of age. In a parallel study, NAC- and CYST-treated and control B/W mice were examined at 24 weeks of age for interval renal histopathology, lymphocyte adhesion molecule expression, and antibody titers and in vitro cytokine production in response to immunization with DNP-KLH. CYST significantly suppressed development of albuminuria and azotemia at 36 and 42 weeks of age compared to control and NAC-treated mice. NAC significantly suppressed anti-DNA antibody levels at 24 weeks. In contrast CYST significantly increased anti-DNA antibody levels at 18 weeks of age (P < 0.001 CYST vs control and NAC-treated mice). Kidneys of CYST-treated mice also had accelerated inflammatory histologic changes despite their lower incidence of albuminuria and azotemia. Mean (+/- s.e.m.) survival of control mice was 33 +/- 2 weeks compared to 38 +/- 2 weeks in NAC-treated mice (P < 0.05 vs control), and 48 +/- 2 weeks in the CYST-treated group (P < 0.01 vs control mice). The antioxidants, NAC and CYST, significantly improved mortality in the female B/W mouse model of SLE. NAC suppressed autoantibody formation and modestly prolonged survival. CYST, despite its augmentation of anti-DNA levels and renal inflammatory changes, inhibited the development of renal insufficiency and markedly improved survival. These findings suggest that ROIs play a role in the pathogenesis of lupus nephritis and that antioxidants reduce the damage causing renal insufficiency. Antioxidants may be a beneficial adjunctive therapy in the treatment of human SLE.

Acetylcysteine↗

Human milk-derived growth factor prevents duodenal ulcer formation.

Human milk was fractionated to obtain a partially purified growth factor preparation. The growth factor in this fraction, designated as human milk growth factor III, exhibits chromatographic and biological characteristics similar to epidermal growth factor-urogastrone. Pretreatment of mice with human milk growth factor III significantly reduces the incidence, number, total length, and severity score of cysteamine-induced duodenal ulcers.

Animals↗

In vitro conditions modify immunoassayability of bovine pituitary prolactin and growth hormone: insights into their secretory granule storage forms.

The amount of immunoassayable intracellular bovine (b) PRL and GH varies depending on treatment conditions. The present studies were designed to characterize the mechanisms involved and to compare immunoassayability of both hormones under similar conditions. Pituitary homogenate and secretory granule hormones displayed both time- and temperature-dependent increases when incubated at pH 10.5 with reduced glutathione; for example, 180 min values were 156% (GH) and 439% (PRL) at 22 C compared with control homogenate values. Under these conditions, PRL in granule cores (the sedimentable fraction after hypotonic lysis of granules) increased the most, changing 120-fold from 2.9 +/- 0.25 micrograms/ml (pH 8.3 control) to 349 +/- 22.8 micrograms/ml. Isolated oligomeric PRL increased significantly after exposure to mercaptoethanol, indicating that changes in assayability do not require nonhormonal granule components. No change in monomeric PRL, or GH in either form, was observed. Pretreatment with iodoacetamide resulted in a 33% decrease in maximal PRL values, indicating the presence and functional importance of granule thiols. Cysteamine inhibition of bPRL values in cores was modulated by urea, EDTA, and iodoacetamide. Though 5 M urea or 0.5% sodium dodecyl sulfate (SDS) increased granule PRL at pH 8.3, maximal values required thiols at pH 10.5. In contrast, maximal GH values were obtained with SDS at pH 8.3 without thiols. Changes in immunoassayability seem to reflect conversion from poorly immunoactive tissue hormone oligomers to monomeric hormone. The data indicate that oligomeric bPRL is stabilized primarily by intermolecular disulfide bonds, although it is also susceptible to urea, SDS, and EDTA; granule thiols may also influence the conversion to monomer. The storage form of bGH appears to be stabilized differently. Maneuvers demonstrated in these studies to influence immunoassayability correlate very well with their previously established effects on hormone release and secretion, strengthening the likelihood that a functional link exists between assayability and secretion.

Animals↗