Serum Paraoxonase, Arylesterase, and Glutathione-S-Transferase Activities and Oxidative Stress Levels in Patients with Mushroom Poisoning.

OBJECTIVES: Consumption of toxic species of mushrooms may have detrimental effects and increase oxidative stress. Paraoxonase, arylesterase and glutathione-S-transferase are antioxidants that resist oxidative stress. In this study, we analyzed the changes in these enzymes during intoxication due to mushrooms. METHODS: The study enrolled 49 adult patients with a diagnosis of mushroom poisoning according to clinical findings and 49 healthy volunteers as the control group. The patients with mild clinical findings were hospitalized due to the possibility that the patient had also eaten the mushrooms and due to clinical findings in the late period, which could be fatal. Paraoxonase, arylesterase, and glutathione-S-transferase concentrations, as well as total antioxidant and oxidant status, were determined in the 49 patients and 49 healthy volunteers by taking blood samples in the emergency department. RESULTS: While paraoxonase, arylesterase, and total antioxidant status were significantly decreased in the patient group (p<0.05), glutathione-S-transferase, total oxidant status and the oxidative stress index were significantly higher (p<0.05). There was a positive correlation between the hospitalization time and the oxidative stress index (r=0.752, p<0.001), whereas a negative correlation was found with glutathione-S-transferase (r=-0.420, p=0.003). CONCLUSION: We observed a significant decrease in paraoxonase and arylesterase and an increase in glutathione-S-transferase and oxidative stress indexes in patients with mushroom poisoning, indicating that these patients had an oxidative status. In particular, a low total antioxidant status and high oxidative stress index may gain importance in terms of the assessment of hospitalization duration.

Serum paraoxonase, arylesterase, and glutathione-s-transferase activities and oxidative stress levels in patients with mushroom poisoning

OBJECTIVES: Consumption of toxic species of mushrooms may have detrimental effects and increase oxidative stress. Paraoxonase, arylesterase and glutathione-S-transferase are antioxidants that resist oxidative stress. In this study, we analyzed the changes in these enzymes during intoxication due to mushrooms. METHODS: The study enrolled 49 adult patients with a diagnosis of mushroom poisoning according to clinical findings and 49 healthy volunteers as the control group. The patients with mild clinical findings were hospitalized due to the possibility that the patient had also eaten the mushrooms and due to clinical findings in the late period, which could be fatal. Paraoxonase, arylesterase, and glutathione-S-transferase concentrations, as well as total antioxidant and oxidant status, were determined in the 49 patients and 49 healthy volunteers by taking blood samples in the emergency department. RESULTS: While paraoxonase, arylesterase, and total antioxidant status were significantly decreased in the patient group (p<0.05), glutathione-S-transferase, total oxidant status and the oxidative stress index were significantly higher (p<0.05). There was a positive correlation between the hospitalization time and the oxidative stress index (r=0.752, p<0.001), whereas a negative correlation was found with glutathione-S-transferase (r=-0.420, p=0.003). CONCLUSION: We observed a significant decrease in paraoxonase and arylesterase and an increase in glutathione-S-transferase and oxidative stress indexes in patients with mushroom poisoning, indicating that these patients had an oxidative status. In particular, a low total antioxidant status and high oxidative stress index may gain importance in terms of the assessment of hospitalization duration.

New in silico insights into the inhibition of RNAP II by α-amanitin and the protective effect mediated by effective antidotes.

Poisonous α-amanitin-containing mushrooms are responsible for the major cases of fatalities after mushroom ingestion. α-Amanitin is known to inhibit the RNA polymerase II (RNAP II), although the underlying mechanisms are not fully understood. Benzylpenicillin, ceftazidime and silybin have been the most frequently used drugs in the management of α-amanitin poisoning, mostly based on empirical rationale. The present study provides an in silico insight into the inhibition of RNAP II by α-amanitin and also on the interaction of the antidotes on the active site of this enzyme. Docking and molecular dynamics (MD) simulations combined with molecular mechanics-generalized Born surface area method (MM-GBSA) were carried out to investigate the binding of α-amanitin and three antidotes benzylpenicillin, ceftazidime and silybin to RNAP II. Our results reveal that α-amanitin should affects RNAP II transcription by compromising trigger loop (TL) function. The observed direct interactions between α-amanitin and TL residues Leu1081, Asn1082, Thr1083, His1085 and Gly1088 alters the elongation process and thus contribute to the inhibition of RNAP II. We also present evidences that α-amanitin can interact directly with the bridge helix residues Gly819, Gly820 and Glu822, and indirectly with His816 and Phe815. This destabilizes the bridge helix, possibly causing RNAP II activity loss. We demonstrate that benzylpenicillin, ceftazidime and silybin are able to bind to the same site as α-amanitin, although not replicating the unique α-amanitin binding mode. They establish considerably less intermolecular interactions and the ones existing are essential confine to the bridge helix and adjacent residues. Therefore, the therapeutic effect of these antidotes does not seem to be directly related with binding to RNAP II. RNAP II α-amanitin binding site can be divided into specific zones with different properties providing a reliable platform for the structure-based drug design of novel antidotes for α-amatoxin poisoning. An ideal drug candidate should be a competitive RNAP II binder that interacts with Arg726, Ile756, Ala759, Gln760 and Gln767, but not with TL and bridge helix residues.

Elevated cardiac enzymes due to mushroom poisoning.

Mushroom poisoning is an important reason of plant toxicity. Wild mushrooms that gathered from pastures and forests can be dangerous for human health. The clinical outcomes and symptoms of mushroom toxicity vary from mild gastrointestinal symptoms to acute multiple organ failure. Toxic effects to kidney and liver of amatoxin are common but cardiotoxic effects are unusual. In this case, we reported the cardiotoxic effect of amatoxin with the elevated troponin-I without any additional finding in electrocardiography, echocardiography and angiography.

Mushroom toxicosis in dogs in general practice causing gastroenteritis, ptyalism and elevated serum lipase activity.

Mushroom toxicosis is rarely diagnosed in dogs and is poorly reported in the veterinary literature. This report suggests that mushroom toxicosis is a potentially under-diagnosed condition in first opinion practice in the UK. Nine dogs with clinical signs consistent with mushroom toxicosis were identified from the records of an out-of-hours emergency service between August 2010 and January 2011. Four dogs were later excluded because of clinical inconsistencies. Clinical signs included acute profuse ptyalism (5/5), diarrhoea (5/5), vomiting (4/5), hypovolaemia (4/5), stuporous (3/5) or obtunded mentation (1/5), miosis (2/5) and hypothermia (2/5). Serum lipase activity was elevated in 4/4 dogs; canine-specific pancreatic lipase was elevated in the remaining dog. Four dogs recovered with aggressive intravenous fluid therapy, analgesia and supportive care; the remaining dog was euthanased due to severe clinical signs and financial constraints. Mushroom toxicosis is an important differential diagnosis for acute gastroenteritis and one possible cause of some cases of "Seasonal Canine Illness". Affected dogs may demonstrate elevated pancreatic enzymes and mushroom toxicosis should be considered in cases of elevated lipase or abnormal semi-quantitative canine-specific pancreatic lipase activities.

[Certain enzymatic markers of liver damage in poisoning with Amanita phalloides]. / Niektóre enzymatyczne wskazniki uszkodzenia watroby w zatruciu muchomorem sromotnikowym.

The activity of selected enzymes was studied in 12 persons on the first day of poisoning with Amanita phalloides. It was found that the most sensitive marker of liver damage was significant increase of activity of cobalt-activated acylase, observed in all studied cases.

Profile of alkaline phosphatase isoenzymes in ten patients poisoned by mushrooms of the genus Lepiota.

Mushrooms of the genus Lepiota (helveola and bruneo-incarnata), similar to those of the genus Amanita, contain amatoxins. Amatoxins, especially amanitin, cause cellular destruction by inhibiting RNA polymerase. Due to the hepatic toxicity of these mushrooms, we have assessed their incidence on alkaline phosphatase levels and on its isoenzymes. Total alkaline phosphatase activity levels were not found to be increased except in two patients, and then only moderately. As regards isoenzymes, the occurrence of a double hepatic fraction in five of the 10 patients, is the most remarkable finding. There seems to exist a relatively close correspondence between the occurrence of a hepatic2 fraction correlating with those of urine amanitin. We conclude that the hepatic2 fraction proves to be important in assessing liver damage by mushroom poisoning because of its correlation with the patient's degree of poisoning.

Effects of picroliv, the active principle of Picrorhiza kurroa, on biochemical changes in rat liver poisoned by Amanita phalloides.

The efficacy of Picroliv, a standardized iridoid glycoside fraction of Picrorhiza kurroa, was studied against the Amanita phalloides-induced biochemical changes in rat liver. A phalloides (50 mg.kg-1) caused significant increases in the activities of hepatic 5'-nucleotidase, gamma-glutamyl transpeptidase, acid ribonuclease, and succinate dehydrogenase, but a decrease in glucose-6-phosphatase. The level of cytochrome P-450 in microsomal fraction and content of glycogen in liver showed significant depletions. Picroliv (25 mg.kg-1.d-1 x 10 d) provided significant restorations of all the biochemical changes poisoned by A phalloides except cytochrome P-450 and glycogen. These results demonstrated the protective effect of Picroliv against A phalloides-induced hepatotoxicity in rats.

[Activity of enzymes of xenobiotic metabolism in the liver of rats with vitamin A deficiency and mycotoxicosis T-2]. / Aktivnost' fermentov metabolizma ksenobiotikov v pecheni krys pri nedostatochnosti vitamina A i mikotoksikoze T-2.

Keeping male rats within a month on a ration deficient in vitamin A led to a distinct decrease in content of cytochrome P-450, in activities of carboxylase, epoxide hydrolase, aniline hydrolase and to a slight inhibition of UDP-glucuronosyl transferase in live tissue. At the same time, activity of glutathione transferase and content of reduced glutathione in liver tissue were increased. After administration of the epoxide-containing T-2 mycotoxin into rats within 10 days at a dose of 0.54 mg/kg activity of the enzymes catalyzing metabolism of xenobiotics was inhibited in the animals maintained on the complete half-synthetic ration, except of epoxide hydrolase and glutathione transferase, activity of which was elevated. The administration of T-2 toxin under conditions of deficiency in vitamin A caused especially distinct inhibition of the enzymes involved in the 1 phase of xenobiotic metabolism but it was accompanied by only slight increase in T-2 toxicosis. The enzymes participating in conjugation of xenobiotics as well as epoxide hydrolase appear to play major roles in detoxication of T-2 mycotoxin.

Clinical findings and follow-up evaluation of an outbreak of mushroom poisoning--survey of Amanita phalloides poisoning.

One hundred and sixty cases of mushroom poisoning during the period July-November 1981 are reported. The survey details 116 observations of short incubation syndromes and 44 cases of delayed syndrome, identified as Amanita Phalloides poisoning. Of the latter, 40 patients were adult (mean age 46 years, range 20-77; 18 females and 22 males) and 4 were children (less than or equal to 12 years old; 3 females and 1 male). All the patients with Amanita Phalloides poisoning were treated according to a therapeutic protocol, based on the infusion of high doses of penicillin G, administration of dexamethasone and thioctic acid, careful correction of water and electrolyte unbalance. The severity of the disease varied in the population of 44 patients: 4 patients died (2 females, 10 and 77 years old; 2 males, 56 and 64 years old); 26 patients were discharged from the hospital as clinically cured; 14 were discharged with persistently abnormal levels of transaminases and they were advised of a follow-up evaluation. The average length of stay in hospital was 2 weeks. Of the patients followed-up, 6 were symptom-free after 6 months, with normal transaminase values and a normal histopathological picture of liver biopsy specimens. In the remaining patients, there was no normalization of transaminase values and liver biopsy specimens showed a picture of chronic active hepatitis. These patients displayed abnormal immunological tests, with presence of immune complexes and of anti-smooth muscle autoantibodies. The results indicate that Amanita Phalloides poisoning represents a threat not only in the high mortality acute phase, but also in the development of chronic active hepatitis in some survivors.

[Treatment of Amanita phalloides poisoning with silybin in combination with penicillin and cortisone]. / Therapie der Knollenblätterpilzvergiftung mit Silibinin in Kombination mit Penicillin und Cortison.

A report is presented of Amanita phalloides poisoning in a family of 3 members admitted with acute gastrointestinal symptoms after eating a meal of various self-picked mushrooms. A latent period of 11 hours preceded the onset of symptoms, a typical feature of this form of poisoning. There was an enormous increase in liver-specific enzymes in two of the cases. The clinical picture was markedly mitigated by the early initiation of silybin therapy, in combination with penicillin and cortisone. The patients made a rapid symptomatic recovery and the liver parameters were sufficiently improved by the 12th day so that the patients could be discharged.

A trend in the therapy of Amanita phalloides poisoning.

Recent experimental evidences have been produced on the protection afforded by penicillin G in rats poisoned by Amanita phalloides extracts. A therapeutic trend which combines penicillin G infusions to the classical supportive measures was applied to 33 cases of severe A. phalloides poisoning, with 100% survival rates. The possible mechanism of the protective effect of penicillin G in A. phalloides poisoning is discussed.