amanitins and amatoxin

Amanita neoovoidea (genus Amanita Pers.) poisoning leads to acute renal failure. Here, we present seven case reports of acute renal failure with acute hepatic failure due to ingestion of A. neoovoidea. Clinical manifestations included gastrointestinal symptoms 1-72 h after ingestion; elevation of renal parameters and blood uric acid, blood urea nitrogen, and creatinine levels; a few abnormal hepatic parameters, primarily albumin decrease and alanine aminotransferase increase; and elevation of zymogram parameters such as cholinesterase and lactate dehydrogenase. To determine whether the hepatic/renal lesions were caused by amanitins, we analyzed the blood and urine samples of patients and specimens of poisonous mushrooms. Morphological and molecular biological analyses indicated that the mushroom was A. neoovoidea. However, no amatoxins and phallotoxins were detected in its basidiomata.

Topics: Acute Kidney Injury; Adult; Aged; Aged, 80 and over; Amanita; Amanitins; Blood Urea Nitrogen; China; Chromatography, High Pressure Liquid; Creatinine; Female; Humans; Male; Middle Aged; Mushroom Poisoning; Uric Acid

Amatoxins are produced primarily by 3 species of mushrooms: Amanita, Lepiota, and Galerina. Because amatoxin poisonings are increasing, the objective of this review was to identify all amatoxin-containing mushroom species, present a toxidromic approach to earlier diagnoses, and compare the efficacies and outcomes of therapies. To meet these objectives, Internet search engines were queried with keywords to select peer-reviewed scientific articles on amatoxin-containing mushroom poisoning and management. Descriptive epidemiological analyses have documented that most mushroom poisonings are caused by unknown mushrooms, and most fatal mushroom poisonings are caused by amatoxin-containing mushrooms. Amanita species cause more fatal mushroom poisonings than other amatoxin-containing species, such as Galerina and Lepiota. Amanita phalloides is responsible for most fatalities, followed by Amanita virosa and Amanita verna. The most frequently reported fatal Lepiota ingestions are due to Lepiota brunneoincarnata, and the most frequently reported fatal Galerina species ingestions are due to Galerina marginata. With the exception of liver transplantation, the current treatment strategies for amatoxin poisoning are all supportive and have not been subjected to rigorous efficacy testing in randomized controlled trials. All patients with symptoms of late-appearing gastrointestinal toxicity with or without false recovery or quiescent periods preceding acute liver insufficiency should be referred to centers providing liver transplantation. Patients with amatoxin-induced acute liver insufficiency that does not progress to liver failure will have a more favorable survival profile with supportive care than patients with amatoxin-induced acute liver failure, about half of whom will require liver transplantation.

Topics: Agaricales; Amanita; Amanitins; Hepatic Insufficiency; Humans; Liver Failure, Acute; Liver Transplantation; Mushroom Poisoning

Amanita phalloides poisoning with a high mortality is a serious health problem in the world. The typical clinical manifestations are usually characterized by the absence of any symptoms followed by severe gastrointestinal disorders and acute liver failure. Inhibition of RNA polymeraseII (RNAP II) activity, apoptosis, and oxidative stress are considered as the major mechanism of amatoxins intoxication. The current treatment measures mainly include prevention of amatoxins absorption, elimination of absorbed amatoxins, potential antidotes therapy, and liver transplantation. Nevertheless, there are no widely accepted treatment criteria for Amanita phalloides poisoning. This paper will focus on the treatment measures based on the previous studies and provide the currently available information for clinicians.

Topics: Acetylcysteine; Amanita; Amanitins; Antidotes; Bile Ducts; Charcoal; Diuresis; Gastric Lavage; Humans; Liver; Liver Failure, Acute; Liver Transplantation; Mushroom Poisoning; Penicillin G; Silymarin

Topics: Acute Disease; Adult; Aged; Amanitins; Female; Glasgow Coma Scale; Hepatic Encephalopathy; Hong Kong; Humans; Liver Transplantation; Male; Middle Aged; Mushroom Poisoning

A shortcut review was carried out to establish whether silibinin is better than conservative management at reducing liver transplantation and death after poisoning with amatoxin-containing mushrooms. Thirty-eight papers were found in Medline and 86 in EMBASE using the reported searches. Of these, five presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results and study weaknesses of these best papers are tabulated. It is concluded that the evidence is limited, but given the lack of alternative treatments in patients with suspected amatoxin-containing mushroom poisoning and the relatively few adverse effects, silibinin should be considered in some patients.

Topics: Amanitins; Antioxidants; Evidence-Based Emergency Medicine; Humans; Mushroom Poisoning; Silybin; Silymarin

Clinical toxicological analysis can significantly contribute toward the confirmation or exclusion of poisoning, especially if clinical signs and symptoms of unknown origin have to be explained. It may be of help when planning specific, but risky, poisoning therapies. Besides frequently used immunoassays for the detection of drugs of abuse, of a small number of medical drugs, and of amatoxins. Chromatographic methods with mass-selective detectors are available in specialized toxicology laboratories. The results of toxicological analyses have to be evaluated and interpreted carefully. Poison control centers can offer support for all medical aspects of poisoning including lab investigations.

Topics: Amanitins; Drug Overdose; Gas Chromatography-Mass Spectrometry; Humans; Illicit Drugs; Immunoassay; Poisoning; Poisons; Prescription Drugs; Toxicology

Topics: Acetylcysteine; Amanita; Amanitins; Bile; Blood Component Removal; Charcoal; Chromatography, High Pressure Liquid; Drainage; Gastric Lavage; Humans; Japan; Laxatives; Liver Transplantation; Mushroom Poisoning; Penicillin G; Silybin; Silymarin

Topics: Acetylcysteine; Adult; Amanita; Amanitins; Cause of Death; Combined Modality Therapy; Critical Care; Diagnosis, Differential; Hepatic Encephalopathy; Humans; Liver Function Tests; Liver Transplantation; Male; Mushroom Poisoning; Prognosis; Silybin; Silymarin

Diagnosis and management of Amanita mushroom poisoning is a challenging problem for physicians across the United States. With 5902 mushroom exposures and two resultant deaths directly linked to Amanita ingestion in 2009, it is difficult for physicians to determine which patients are at risk for lethal toxicity. Identification of amatoxin poisoning can prove to be difficult due to delay in onset of symptoms and difficulty with identification of mushrooms. Consequently, it is difficult for the Emergency Physician to determine proper disposition. Further, treatment options are controversial.. To review current data to help health care providers effectively identify and treat potentially deadly Amanita mushroom ingestions.. We present two cases of Amanita mushroom ingestion in the northeastern United States treated with N-acetylcysteine, high-dose penicillin, cimetidine, and silibinin, a semi-purified fraction of milk thistle-derived silymarin, as part of their treatment regimen. The mushroom species was identified by a consultant as Amanita Ocreata.. We present the successful treatment of 2 patients who ingested what we believe to be an Amanita species never before identified in the northeastern United States.

Topics: Aged; Amanita; Amanitins; Antioxidants; Female; Humans; Male; Middle Aged; Mushroom Poisoning; Treatment Outcome

More than 90% of all fatal mushroom poisonings worldwide are due to amatoxin containing species that grow abundantly in Europe, South Asia, and the Indian subcontinent. Many cases have also been reported in North America. Initial symptoms of abdominal cramps, vomiting, and a severe cholera-like diarrhea generally do not manifest until at least six to eight hours following ingestion and can be followed by renal and hepatic failure. Outcomes range from complete recovery to fulminant organ failure and death which can sometimes be averted by liver transplant. There are no controlled clinical studies available due to ethical reasons, but uncontrolled trials and case reports describe successful treatment with intravenous silibinin (Legalon® SIL). In nearly 1,500 documented cases, the overall mortality in patients treated with Legalon® SIL is less than 10% in comparison to more than 20% when using penicillin or a combination of silibinin and penicillin. Silibinin, a proven antioxidative and anti-inflammatory acting flavonolignan isolated from milk thistle extracts, has been shown to interact with specific hepatic transport proteins blocking cellular amatoxin re-uptake and thus interrupting enterohepatic circulation of the toxin. The addition of intravenous silibinin to aggressive intravenous fluid management serves to arrest and allow reversal of the manifestation of fulminant hepatic failure, even in severely poisoned patients. These findings together with the available clinical experience justify the use of silibinin as Legalon® SIL in Amanita poisoning cases.

Topics: Amanitins; Animals; Antidotes; Chemical and Drug Induced Liver Injury; Humans; Molecular Structure; Mushroom Poisoning; Silymarin; Tissue Distribution; Treatment Outcome

Topics: Agaricales; Amanitins; Cyanides; Humans; Japan; Mushroom Poisoning; Peptides, Cyclic

The tryptathionine linkage is a crosslink formed between tryptophan and cysteine. This feature is characteristic of the bicyclic peptides: the phallotoxins and the amatoxins. These peptides both bind to protein folds of their respective targets (F-actin and RNA pol II, respectively) with extremely high affinities. Studies on these peptides have shown that the tryptathionine crosslink is essential for this binding affinity. Tryptathionines have been investigated for many years and several syntheses exist for their formation. In this review, we report on the various methodologies employed in tryptathionine synthesis, and discuss some of the advantages and disadvantages associated with each of them.

Topics: Amanitins; Cross-Linking Reagents; Cysteine; Electrons; Indoles; Peptides; Tryptophan

Mushrooms are ubiquitous in nature. They are an important source of nutrition; however, certain varieties contain chemicals that can be highly toxic to humans. Industrially cultivated mushrooms are historically very safe, but foraging for mushrooms or accidental ingestion of mushrooms in the environment can result in serious illness and death. The emergency department is the most common site of presentation for patients suffering from acute mushroom poisoning. Although recognition can be facilitated by identification of a characteristic toxidrome, the presenting manifestations can be variable and have considerable overlap with more common and generally benign clinical syndromes. The goal of this two-part article is to review the knowledge base on this subject and provide information that will assist the clinician in the early consideration, diagnosis and treatment of mushroom poisoning. Part I, presented in this issue of the Journal, reviews the epidemiology and demographics of mushroom poisoning, the physical characteristics of the most toxic varieties, the classification of the toxic species, and an overview of the cyclopeptide-containing mushroom class. Part II, to be published in the next issue of the Journal, will be focused on the presentation of the other classes of toxic mushrooms along with an up-to-date review of the most recently identified poisonous varieties.

Topics: Agaricales; Amanita; Amanitins; Child, Preschool; Humans; Mushroom Poisoning; Mycotoxins

Among fungal toxins causing organ damage in the human body, amatoxins and orellanine remain exceptional. Amatoxins, a group of bicyclic octapeptides occurring in some Amanita, Galerina and Lepiota species, induce deficient protein synthesis resulting in cell death, but might also exert toxicity through inducing apoptosis. Target organs are intestinal mucosa, liver and kidneys. Poisoning will result in dehydration and electrolyte derangement, liver necrosis and possibly kidney damage. In established poisoning the mainstay of treatment is optimum symptomatic and supportive care. No specific treatment is available, but some pharmaceuticals, like silibinin, benzylpenicillin and acetylcysteine, might have a role in limiting the extent of hepatic damage. Orellanine is a nephrotoxic bipyridine N-oxide found in some Cortinarius species. Its mechanism of action is not fully understood, but it has been shown to inhibit protein synthesis and to generate free oxygen radicals. As early symptoms often are lacking or vague, poisoning may initially be overlooked or misinterpreted and the patients usually present with established renal damage. Supportive care is the only therapeutic option. Tricholoma equestre might contain a myotoxin and repeated ingestion may cause significant rhabdomyolysis. Ingestion of Amanita smithiana and A. proxima has been reported to result in kidney damage. Gyromitrin, a toxic compound that is converted to hydrazines in the stomach, occurs in some Gyromitra species. It is mainly neurotoxic, but may also induce moderate hepatic damage and haemolysis.

Topics: 2,2'-Dipyridyl; Acetaldehyde; Amanitins; Basidiomycota; Humans; Kidney Diseases; Mushroom Poisoning; Mycotoxins; Rhabdomyolysis

Amatoxin poisoning is a medical emergency characterized by a long incubation time lag, gastrointestinal and hepatotoxic phases, coma, and death. This mushroom intoxication is ascribed to 35 amatoxin-containing species belonging to three genera: Amanita, Galerina, and Lepiota. The major amatoxins, the alpha-, beta-, and gamma-amanitins, are bicyclic octapeptide derivatives that damage the liver and kidney via irreversible binding to RNA polymerase II.. The mycology and clinical syndrome of amatoxin poisoning are reviewed. Clinical data from 2108 hospitalized amatoxin poisoning exposures as reported in the medical literature from North America and Europe over the last 20 years were compiled. Preliminary medical care, supportive measures, specific treatments used singly or in combination, and liver transplantation were characterized. Specific treatments consisted of detoxication procedures (e.g., toxin removal from bile and urine, and extracorporeal purification) and administration of drugs. Chemotherapy included benzylpenicillin or other beta-lactam antibiotics, silymarin complex, thioctic acid, antioxidant drugs, hormones and steroids administered singly, or more usually, in combination. Supportive measures alone and 10 specific treatment regimens were analyzed relative to mortality.. Benzylpenicillin (Penicillin G) alone and in association was the mostfrequently utilized chemotherapy but showed little efficacy. No benefit was found for the use of thioctic acid or steroids. Chi-square statistical comparison of survivors and dead vs. treated individuals supported silybin, administered either as mono-chemotherapy or in drug combination and N-acetylcysteine as mono-chemotherapy as the most effective therapeutic modes. Future clinical research should focus on confirming the efficacy of silybin, N-acetylcysteine, and detoxication procedures.

Topics: Agaricales; Amanitins; Animals; Chemical and Drug Induced Liver Injury; Emergency Medical Services; Humans; Liver Transplantation; Mushroom Poisoning; Retrospective Studies

Amanita toxic peptides are the main toxins in wild mushrooms and often result in animal and human intoxication and death. Due to the specific inhibition to eukaryotic cell RNA polymerases II by amatoxins and the binding to actin by phallotoxins, amanita toxic peptides play important roles in life science researches. This paper presents a latest review of applications of amanita toxic peptides in life scientific researches.

Topics: Amanita; Amanitins; Animals; DNA-Directed RNA Polymerases; Fungal Proteins; Humans

The most poisonous mushroom toxins are produced by Amanita phalloides (death cap). The occurrence and chemistry of three groups of toxins (amatoxins, phallotoxins and virotoxins) are summarized. The concentration and distribution of toxins in certain species are variable, with the young fruit body containing lower, and the well-developed fungus higher concentrations, but there is a high variability among specimens collected in the same region. Regarding phallotoxins, the volva (the ring) is the most poisonous. The most important biochemical effect of amatoxins is the inhibition of RNA polymerases (especially polymerase II). This interaction leads to a tight complex and the inhibition is of a non-competitive type. Non-mammalian polymerases show little sensitivity to amanitins. The amatoxins cause necrosis of the liver, also partly in the kidney, with the cellular changes causing the fragmentation and segregation of all nuclear components. Various groups of somatic cells of emanation resistance have been isolated, including from a mutant of Drosophila melanogaster. The phallotoxins stimulate the polymerization of G-actin and stabilize the F-actin filaments. The interaction of phallotoxins occurs via the small, 15-membered ring, on the left side of the spatial formula. The symptoms of human poisoning and the changes in toxin concentrations in different organs are summarized. Conventional therapy includes: (1) stabilization of patient's condition with the correction of hypoglycaemia and electrolytes; (2) decontamination; and (3) chemotherapy with different compounds. Finally, certain antagonists and protective compounds are reviewed, bearing in mind that today these have more of a theoretical than a practical role.

Topics: Amanita; Amanitins; Animals; Humans; Models, Molecular; Molecular Structure; Mushroom Poisoning; Peptides, Cyclic

Methods for the chromatographic determination of amanitins, toxins of Amanita phalloides (Fr.), Link mushrooms and related toxins are reviewed; particular emphasis is given to high-performance liquid chromatographic methods. The main chemical and toxicological aspects are discussed, but the focus of the present review is on the analytical problems arising in a laboratory charged with the setting up of a procedure which can direct the appropriate clinical management of an intoxicated patient or solve a forensic case.

Topics: Amanita; Amanitins; Chromatography; Humans; Mycotoxins

Hepatotoxic mushroom poisoning (due to Amanita, Lepiota and Galerina species) may be considered as a real medical emergency, since an early diagnosis and immediate treatment are required for a successful outcome. In this review the physio-pathological features and the clinical picture of amatoxin poisonings are described as the basis for diagnosis and therapeutic decisions. The treatment schedule proposed is analyzed in some points: Symptomatic and supportive measures, toxin removal and extraction procedures, and the possibility of using antidotes. Some parameters with prognostic significance are commented on. Finally, the mortality rate and its evolution throughout the present century is also considered.

Topics: Amanitins; Combined Modality Therapy; Humans; Mushroom Poisoning; Prognosis

This review gives a comprehensive account of the molecular toxicology of the bicyclic peptides obtained from the poisonous mushrooms of the genus Amanita. The discussion of the biochemical events will be preceded by a consideration of the chemistry of the toxic peptides. The structural features essential for biological activities of both the amatoxins and the phallotoxins will be discussed, also including the most important analytical data. Similar consideration will be given to antamanide, a cyclic peptide, which counteracts phalloidin. In addition, the phallolysins, three cytolytic proteins from Amanita phalloides will be discussed. The report on the biological activity of the amatoxins will deal with the sensitivity of the different RNA-polymerases towards the toxins and with their action on various cell types. Consideration will also be given to systems in which alpha-amanitin was used and can be used as a molecular tool; in the past, many investigators used the inhibitor in molecular biology, genetics, and even in physiological research. As for the phallotoxins, discussion of the affinity of these toxins for actin is provied. Further discussion attempts to understand the course of intoxication by filling in the gap between the first molecular event, formation of microfilaments, and the various lesions in hepatocytes during the intoxication.

Topics: Agaricales; Amanita; Amanitins; Animals; DNA-Directed RNA Polymerases; Erythrocytes; Humans; Mycotoxins; Peptides, Cyclic; Protein Binding; Protein Conformation; RNA Polymerase II; Species Specificity; Transcription, Genetic

A 60-year-old man presented with acute gastroenteritis, hypovolemic shock, acute renal failure (BUN/Cr, 56.7/4.24 mg/dl), and aspiration pneumonia. The previous day, he ingested 30 caps of mushrooms of an unknown species. The patient was treated with a massive intravenous infusion, renal replacement therapy, and antimicrobial agents. Late-onset mild liver injury peaked on day 11 (AST/ALT, 62/67 IU/l). Acute renal failure improved once before worsening, with the worst symptoms on day 19 (BUN/Cr, 99/6.61 mg/dl). Thereafter, the patient showed gradual improvement, and renal replacement therapy was discontinued on day 23. His general condition improved fully and he was transferred to another hospital for rehabilitation on day 47. The mushrooms were later identified as Galerina sulciceps by the Basic Local Alignment Search Tool, and toxicologic analysis using liquid chromatography-tandem mass spectrometry revealed an average of 85 ppm α-amanitin and 330 ppm β-amanitin in the tissue of the mushrooms brought in by the patient's family. Galerina sulciceps is distributed mainly in tropical and subtropical regions of Southeast Asia and had never been identified before in Japan. The heat of fermentation generated by the thick layer of wood chips on the ground or global warming may have contributed to its growth in Japan. Interestingly, our patient did not have liver dysfunction, which is one main and typical amatoxin poisoning symptom. Variation in clinical presentation may be attributed to the different ratios of α-amanitin to β-amanitin in different mushroom species.

Topics: Acute Kidney Injury; Agaricales; Alpha-Amanitin; Amanitins; Humans; Japan; Male; Middle Aged; Mushroom Poisoning

About 95% of fatal mushroom poisonings worldwide are caused by amatoxins and phallotoxins mostly produced by species of Amanita, Galerina, and Lepiota. The genus Lepiota is supposed to include a high number of species producing amatoxins. In this study, we investigated 16 species of Lepiota based on 48 recently collected specimens for the presence of amatoxins by liquid chromatography coupled to a diode-array detector and mass spectrometry (UHPLC-QTOF-MS/MS). By comparing the retention times, UV absorptions, and diagnostic MS fragment ions with data obtained from the benchmark species Amanita phalloides, we detected α-amanitin and γ-amanitin in Lepiota subincarnata, α-amanitin and amaninamide in Lepiota brunneoincarnata, and β-amanitin and α-amanitin in Lepiota elaiophylla. Phallotoxins have not been detected any of these species. Two possibly undescribed amatoxin derivatives were found in Lepiota boudieri and L. elaiophylla, as well as one further non-amatoxin compound in one specimen of L. cf. boudieri. These compounds might be used to differentiate L. elaiophylla from L. xanthophylla and species within the L. boudieri species complex. No amatoxins were detected in L. aspera, L. castanea, L. clypeolaria, L. cristata, L. erminea, L. felina, L. fuscovinacea, L. lilacea, L. magnispora, L. oreadiformis, L. pseudolilacea, L. sp. (SeSa 5), and L. subalba. By combining the occurrence data of amatoxins with a phylogenetic analysis, a monophyletic group of amatoxin containing species of Lepiota is evident. These chemotaxonomic results highlight the relevance of systematic relationships for the occurrence of amatoxins and expand our knowledge about the toxicity of species of Lepiota.

Topics: Agaricales; Amanitins; Mushroom Poisoning; Phylogeny; Tandem Mass Spectrometry

Amanitin is used extensively as a research tool to inhibit RNA Pol II thereby implicating its role in mRNA transcription. Recently, amanitin has gained traction as a toxic payload for targeted therapy. Here we report the first-ever photocaged amanitin analog, that is non-toxic and can be pre-loaded into cells. Light provides a means to inhibit RNA Pol II and provoke cell death on-demand.

Topics: Amanitins; Animals; Cell Death; Cell Survival; CHO Cells; Cricetulus; Dose-Response Relationship, Drug; Molecular Structure; Photochemical Processes; Prodrugs; RNA Polymerase II

Topics: Acetylcysteine; Amanitins; Humans; Mushroom Poisoning

Topics: Acetylcysteine; Amanitins; Humans; Mushroom Poisoning

Mushroom toxicity is the main branch of foodborne poisoning, and liver damage caused by amatoxin poisoning accounts for more than 90 % of deaths due to mushroom poisoning. Alpha-amatoxin (α-AMA) has been considered the primary toxin from amatoxin-containing mushrooms, which is responsible for hepatotoxicity and death. However, the mechanism underlying liver failure due to α-AMA remains unclear. This study constructed animal and cell models. In the animal experiments, we investigated liver injury in BALB/c mice at different time points after α-AMA treatment, and explored the process of inflammatory infiltration using immunohistochemistry and western blotting. Then, a metabonomics method based on gas chromatography mass spectrometry (GCMS) was established to study the effect of α-AMA on liver metabonomics. The results showed a significant difference in liver metabolism between the exposed and control mice groups that coincided with pathological and biochemical indicators. Moreover, 20 metabolites and 4 metabolic pathways related to its mechanism of action were identified, which suggested that energy disorders related to mitochondrial dysfunction may be one of the causes of death. The significant changes of trehalose and the fluctuation of LC3-II and sqstm1 p62 protein levels indicated that autophagy was also involved in the damage process, suggesting that autophagy may participate in the clearance process of damaged mitochondria after poisoning. Then, we constructed an α-AMA-induced human normal liver cells (L-02 cells) injury model. The above hypothesis was further verified by detecting cell necrosis, mitochondrial reactive oxygen species (mtROS), mitochondrial permeability transition pore (mPTP) opening, mitochondrial membrane potential (Δψ m), and cellular ATP level. Collectively, our results serve as direct evidence of elevated in vivo hepatic mitochondrial metabolism in α-AMA-exposed mice and suggest that mitochondrial dysfunction plays an important role in the early stage of α-AMA induced liver failure.

Topics: Amanitins; Animals; Cell Line; Chemical and Drug Induced Liver Injury; Disease Models, Animal; Energy Metabolism; Humans; Liver; Liver Failure; Metabolomics; Mice, Inbred BALB C; Mitochondria, Liver; Mushroom Poisoning; Time Factors

Topics: Acetylcysteine; Amanitins; Humans; Mushroom Poisoning

Topics: Acetylcysteine; Amanitins; Humans; Mushroom Poisoning

Topics: Amanita; Amanitins; Chromatography, High Pressure Liquid; Cooking; Forensic Toxicology; Hot Temperature; Humans; Mushroom Poisoning

To describe the clinical course, treatment, and outcome of 5 dogs following ingestion of toxic Amanita spp. mushrooms containing amatoxins using an adapted version of the Santa Cruz protocol developed for people.. Five dogs were presented with clinical signs compatible with amanitin toxicity with witnessed ingestion noted in 3 of 5 dogs. Clinical findings included acute onset vomiting and diarrhea, lethargy, and hepatopathy including signs of fulminant hepatic failure (increased liver enzyme activities, hyperbilirubinemia, prolonged clotting times, and hypoglycemia were noted among these cases). Urine toxicological screening confirmed the presence of Amanita toxins in 4 cases with expert mycologist speciation in the fifth. Core interventions included percutaneous biliary drainage, use of octreotide, and early nil per os orders. All dogs survived to discharge with this treatment strategy.. This case series describes the use of a modified version of the Santa Cruz protocol to address amatoxin-induced fulminant hepatic failure in dogs. The protocol was safe, well tolerated, and all patients made a full clinical recovery.

Topics: Amanita; Amanitins; Animals; Dog Diseases; Dogs; Humans; Liver Failure, Acute; Male; Mushroom Poisoning

Topics: Amanitins; Breast Feeding; Female; Humans; Infant; Milk, Human

Some but not all of the species of 'little brown mushrooms' in the genus Galerina contain deadly amatoxins at concentrations equaling those in the death cap, Amanita phalloides. However, Galerina's ~300 species are notoriously difficult to identify by morphology, and the identity of toxin-containing specimens has not been verified with DNA barcode sequencing. This left open the question of which Galerina species contain toxins and which do not. We selected specimens for toxin analysis using a preliminary phylogeny of the fungal DNA barcode region, the ribosomal internal transcribed spacer (ITS) region. Using liquid chromatography/mass spectrometry, we analyzed amatoxins from 70 samples of Galerina and close relatives, collected in western British Columbia, Canada. To put the presence of toxins into a phylogenetic context, we included the 70 samples in maximum likelihood analyses of 438 taxa, using ITS, RNA polymerase II second largest subunit gene (RPB2), and nuclear large subunit ribosomal RNA (LSU) gene sequences. We sequenced barcode DNA from types where possible to aid with applications of names. We detected amatoxins only in the 24 samples of the G. marginata s.l. complex in the Naucoriopsis clade. We delimited 56 putative Galerina species using Automatic Barcode Gap Detection software. Phylogenetic analysis showed moderate to strong support for Galerina infrageneric clades Naucoriopsis, Galerina, Tubariopsis, and Sideroides. Mycenopsis appeared paraphyletic and included Gymnopilus. Amatoxins were not detected in 46 samples from Galerina clades outside of Naucoriopsis or from outgroups. Our data show significant quantities of toxin in all mushrooms tested from the G. marginata s.l. complex. DNA barcoding revealed consistent accuracy in morphology-based identification of specimens to G. marginata s.l. complex. Prompt and careful morphological identification of ingested G. marginata s.l. has the potential to improve patient outcomes by leading to fast and appropriate treatment.

Topics: Agaricales; Amanitins; Humans; Likelihood Functions; Phylogeny

Here we report a scalable synthesis of the key amino acid residue, (2

Topics: Alpha-Amanitin; Amanitins; Amino Acids; Immunoconjugates

Amatoxin poisoning is the main cause of death from accidental ingestion of poisonous mushrooms and a mortality rate of 27.3% has been reported in Thailand. Symptoms of mushroom ingestion are often confused with food poisoning; thus, gastroenteritis is not recognised as the first phase of poisoning. Our study assessed the efficacy of N-acetylcysteine (NAC) as a treatment for amatoxin poisoning. We retrospectively analysed 74 medical records over 12 years. The majority (70/74) were treated successfully with NAC; death in the remaining 4 (5.4%) patients was attributed to late presentation in three and advanced alcoholic cirrhosis in one.

Topics: Acetylcysteine; Amanitins; Female; Gastroenteritis; Humans; Male; Mushroom Poisoning; Retrospective Studies; Thailand; Treatment Outcome

Topics: Amanita; Amanitins; Antioxidants; China; Female; Humans; Liver; Liver Failure, Acute; Male; Middle Aged; Mushroom Poisoning; Thrombocytopenia

Globally, mushroom poisonings cause about 100 human deaths each year, with thousands of people requiring medical assistance. Dogs are also susceptible to mushroom poisonings and require medical assistance. Cyclopeptides, and more specifically amanitins (or amatoxins, here), are the mushroom poison that causes the majority of these deaths. Current methods (predominantly chromatographic, as well as antibody-based) of detecting amatoxins are time-consuming and require expensive equipment. In this work, we demonstrate the utility of the lateral flow immunoassay (LFIA) for the rapid detection of amatoxins in urine samples. The LFIA detects as little as 10 ng/mL of α-amanitin (α-AMA) or γ-AMA, and 100 ng/mL of β-AMA in urine matrices. To demonstrate application of this LFIA for urine analysis, this study examined fortified human urine samples and urine collected from exposed dogs. Urine is sampled directly without the need for any pretreatment, detection from urine is completed in 10 min, and the results are read by eye, without the need for specialized equipment. Analysis of both fortified human urine samples and urine samples collected from intoxicated dogs using the LFIA correlated well with liquid chromatography-mass spectrometry (LC-MS) methods.

Topics: Amanitins; Animals; Dog Diseases; Dogs; Humans; Immunoassay; Molecular Structure; Mushroom Poisoning; Point-of-Care Testing; Sensitivity and Specificity

The mushroom poison that causes the most deaths is the class of toxins known as amatoxins. Current methods to sensitively and selectively detect these toxins are limited by the need for expensive equipment, or they lack accuracy due to cross-reactivity with other chemicals found in mushrooms. In this work, we report the development of a competition-based lateral flow immunoassay (LFIA) for the rapid, portable, selective, and sensitive detection of amatoxins. Our assay clearly indicates the presence of 10 ng/mL of α-AMA or γ-AMA and the method including extraction and detection can be completed in approximately 10 minutes. The test can be easily read by eye and has a presumed shelf-life of at least 1 year. From testing 110 wild mushrooms, the LFIA identified 6 out of 6 species that were known to contain amatoxins. Other poisonous mushrooms known not to contain amatoxins tested negative by LFIA. This LFIA can be used to quickly identify amatoxin-containing mushrooms.

Topics: Amanita; Amanitins; Antibodies; Gold; Immunoassay; Peptides; Reference Standards

In order to explore the role of apoptosis in alpha-amatoxin (α-AMA) induced liver injury and probable upstream activation signals, we established animal and cellular models, respectively, for this pathophysiological condition. To this end, we evaluated the survival rate and serum biochemical parameters in BALB/c mice exposed to α-AMA at different time periods, along with the levels of oxidative and antioxidant enzymes in the liver tissue of these mice and proteins involved in apoptosis-related pathways. Our results reveal that α-AMA-induced apoptosis occurs primarily through the mitochondrial apoptotic pathway and is associated with oxidative damage. Further, in order to verify the key nodes and important upstream activators in this apoptotic pathway, we estimated the levels of p53 protein and downstream mitochondrial apoptotic pathway-related proteins in L-02 cells, all of which were found to change significantly. We also found that the levels of total and mitochondrial reactive oxygen species (ROS) in L-02 cells increased with time. Collectively, our findings suggest that α-AMA affects many cellular processes, including the expression of p53 independent of transcription and the expression of Bax and Bcl-2, thereby activating the subsequent caspase cascade pathways. In addition, we identified ROS to be an upstream signaling molecule involved in the α-AMA-induced apoptosis of mouse liver cells and L-02 cells.

Topics: Amanitins; Animals; Apoptosis; Blotting, Western; Cell Death; Cell Line; Flow Cytometry; In Situ Nick-End Labeling; Liver; Male; Mice; Mice, Inbred BALB C; Mitochondria; Oxidative Stress; Peptides, Cyclic; Reactive Oxygen Species

In this study, we present the preparation of a new reverse-phase/phenylboronic-acid (RP/PBA)-type mixed-mode magnetic solid-phase extraction (MSPE) adsorbent for use in the cleanup of amatoxin- and phallotoxin-containing samples intended for ultrahigh-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) analysis. Further, the RP/PBA magnetic microspheres have phenyl and phenylboronic acid groups on their surfaces that selectively adsorb amatoxins and phallotoxins through hydrophobic, π-π, and boronate affinity, significantly reducing matrix effects in UPLC-MS/MS analysis. After systematic optimization, all the standard calibration curves expressed satisfactory linearity (r > 0.9930), limits of detection (0.3 μg/kg), and recovery (97.6%-114.2%). Compared with other reported methods, this method also has the advantages of simple, fast, and efficient operation using relatively small amounts of the MSPE adsorbent. Furthermore, the method was successfully applied in a poisoning incident caused by Lepiota brunneoincarnata Chodat & C. Martín ingestion.

Topics: Adsorption; Amanitins; Boronic Acids; Chromatography, High Pressure Liquid; Chromatography, Reverse-Phase; Limit of Detection; Magnets; Microspheres; Solid Phase Extraction; Tandem Mass Spectrometry

Amatoxins, most of which are hepatotoxic, can cause fatal intoxication. While mushrooms in the amatoxin-containing Galerina genus are rare, they can poison humans and animals worldwide. Few studies have profiled the toxicity of Galerina marginata. In addition, many studies indicate that macrofungi can have different characteristics in different regions. In this study, the quantities of toxins present in G. marginata from different provinces in Turkey were analysed using reversed-phase high-performance liquid chromatography with ultraviolet detection (RP-HPLC-UV) and liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS). G. marginata samples were collected from three different regions of Turkey. The taxonomic categorization of mushrooms was based on their micro- and macroscopic characteristics. The presence of toxins α-amanitin (AA), β-amanitin (BA), γ-amanitin (GA), phalloidin (PHD) and phallacidin (PHC) quantities were measured using RP-HPLC-UV and then were confirmed using LC-ESI-MS/MS. BA levels were higher than AA levels in G. marginata mushrooms collected from all three regions. Moreover, the levels of GA were below the detection limit and no phallotoxins were detected. This is the first study to identify and test the toxicity of G. marginata collected from three different regions of Turkey using RP-HPLC-UV. This is also the first study to confirm the UV absorption of amatoxins in G. marginata using LC-ESI-MS/MS, which is a far more sensitive process. More studies evaluating the toxicity of G. marginata in other geographic regions of the world are needed.

Topics: Alpha-Amanitin; Amanitins; Mushroom Poisoning; Toxins, Biological; Turkey

A 34-year-old woman from Thai origin developed acute liver failure after ingestion of a soup which contained the death cap (Amanita phalloides).. In patients with poisoning due to amatoxin-containing mushrooms, gastro-intestinal complaints usually develop several hours after ingestion, followed by acute hepatic failure which occasionally leads to death. The incidence of reported mushroom poisonings in the Netherlands has increased in 2019, which is possibly associated with migration of asylum seekers who regularly pick and eat mushrooms.. In the Netherlands mushroom intoxication is rare. Therefore, there is a lack of knowledge among health care personnel and foragers. The present case report highlights the importance of awareness of the poisonous death cap to prevent intoxications and optimize treatment decisions.

Topics: Adult; Amanita; Amanitins; Female; Humans; Liver Failure, Acute; Mushroom Poisoning; Netherlands

Topics: Adult; Amanita; Amanitins; Breast Feeding; Female; Humans; Infant, Newborn; Middle Aged; Milk, Human; Mushroom Poisoning; Treatment Outcome

Eating mushrooms known to contain amatoxin is fraught with serious complications. The analysis of the relevant literature publications revealed no article with the description of the histological picture of the internal organs in the subjects intoxicated with amatoxin. It is known, however, that such poisoning is associated with the severe irreversible injuries to all intracellular protein structures the character of which depends on time. Specifically, acute amatoxin intoxication produces the well apparent clinical picture within 6 days after intake of the poison. It is characterized by acute renal and hepatic insufficiency in the combination with the injury to the conducting system of heart and the myocrardium itself. Thereafter, the disseminated intravascular coagulation (DIC) syndrome developed accompanied by the signs of progressive tissue hypoxia that ended in death on day 9. The histological study has demonstrated necrotic foci in the liver and oedematous hepatic stroma. Kidneys underwent multiple hemorrhages, necrosis of convoluted tubules and well apparent hydropic protein dystrophy of their epithelium. The adrenal glands showed up signs of necrosis and hemorrhage. It is concluded that poisoning with mushrooms (amatoxin) should be regarded as the most probable cause of the condition requiring differential diagnostics between acute gastroenteritis and renal insufficiency.. Употребление в пищу грибов, содержащих аматоксин, опасно серьезными осложнениями. Анализ источников литературы не выявил ни одного сообщения, в котором бы приводилась гистологическая картина внутренних органов у лиц, отравившихся аматоксином. Известно, что при таких отравлениях необратимые изменения обусловлены повреждением всех клеточных белковых структур и имеют различную временнýю составляющую. В случае острого отравления аматоксином развернутую клиническую картину в виде острой почечной и печеночной недостаточности, а также поражения проводящей системы сердца и самого миокарда наблюдали на 6-е сутки. Смерть наступила на 9-е сутки на фоне массивного ДВС-синдрома и признаков прогрессирующей тканевой гипоксии. При гистологическом исследовании наблюдали очаги некроза и отек стромы печени, множественные кровоизлияния, некроз извитых канальцев почек, резко выраженную белковую гидропическую дистрофию эпителия извитых канальцев, некроз и кровоизлияния в ткани надпочечников. Отравление грибами (аматоксином) необходимо рассматривать в качестве наиболее вероятной причины при дифференциальной диагностике острого гастроэнтерита и почечной недостаточности.

Topics: Amanita; Amanitins; Humans; Kidney; Liver; Mushroom Poisoning

Amatoxins (AMAs) are lethal toxins found in a variety of mushroom species. Detection methods are needed to determine the occurrence of AMAs in mushroom species suspected in mushroom poisonings. In this manuscript, we report the generation of novel monoclonal antibodies (mAbs, AMA9G3 and AMA9C12) and the development of a competitive, enzyme-linked immunosorbent assay (cELISA) that is sensitive at 1 ng mL

Topics: Amanita; Amanitins; Animals; Antibodies, Monoclonal; Enzyme-Linked Immunosorbent Assay; Female; Hemocyanins; Mice, Inbred BALB C; Periodic Acid

Mushroom poisoning with Amanita phalloides or similar species can lead to liver failure with 10-30% mortality rates. We aimed at defining the prognostic value of urinary amatoxin quantification in patients with hepatotoxic mushroom poisoning.. Data from 32 patients with hepatotoxic mushroom poisoning (Hospital Clínic Barcelona, 2002-16) in whom urinary amatoxins were determined (ELISA) were retrospectively reviewed. Correlations between urinary amatoxin and collected baseline variables with outcomes including hepatotoxicity (ALT>1000 U/L), severe acute liver injury (ALI, prothrombin <50%), acute liver failure (ALF, ALI and encephalopathy), transplantation/death and hospital length-of-stay, were evaluated.. 12/32 patients developed increased aminotransferase activity. Among the 13/32 amatoxin negative patients, 1 developed ALI and 12/13 no hepatotoxicity. Among the 19/32 amatoxin positive patients, 8/19 (42%) developed hepatotoxicity, including 5 who progressed to severe ALI, of whom 3 developed ALF (2 deaths, 1 transplantation). Urinary amatoxin and prothrombin were independent predictors of hepatotoxicity, ALT peak values (along with age) and hospital length-of-stay. In positive amatoxins patients, urinary concentrations > 55 ng/ml (or a baseline prothrombin ≤ 83%), were associated to hepatotoxicity (presented by 8/9 patients with ALT>1000 U/L). Among 5 patients with urinary amatoxin ≥ 70 ng/ml, 4 developed severe ALI.. In patients with hepatotoxic mushroom poisoning, a negative urinary amatoxin quantification within 72h of intake ruled out the risk of hepatotoxicity in 92% of patients, whereas positive urinary amatoxins were associated with hepatotoxicity and severe ALI. Concentrations >55 ng/ml and ≥ 70 ng/ml were predictive of hepatotoxicity and severe ALI, respectively.

Topics: Adolescent; Adult; Aged; Amanitins; Child; Enzyme-Linked Immunosorbent Assay; Female; Humans; Liver Failure, Acute; Logistic Models; Male; Middle Aged; Mushroom Poisoning; Predictive Value of Tests; Retrospective Studies; Spain; Young Adult

The most frequently reported fatal Lepiota ingestions are due to L. brunneoincarnata. We present a case of L. brunneoincarnata poisoning with endoscopic nasobiliary drainage known to be the first in China. The patient suffered gastrointestinal symptoms 9 h post ingestion of mushrooms. The patient was hospitalized 4 days after eating the mushrooms with jaundice. The peak ALT, AST, APTT, TBIL and DBIL values of the patient were as follow: ALT, 2980 U/L (day 4 post ingestion); AST, 1910 U/L (day 4 post ingestion); APTT, 92.8 seconds (day 8 post ingestion), TBIL, 136 μmol/L (day 10 post ingestion), DBIL 74 μmol/L (day 10 post ingestion). UPLC-ESI-MS/MS was used to detect the peptide toxins in the mushroom and biological samples from the patient. We calculated that the patient may have ingested a total of 29.05 mg amatoxin from 300 g mushrooms, consisting of 19.91 mg α-amanitin, 9.1 mg β-amanitin, and 0.044 mg γ-amanitin. Amatoxins could be detected in bile even on day 6 after ingestion of L. brunneoincarnata. With rehydration, endoscopic nasobiliary drainage and intravenous infusion of Legalon SIL, the patient recovered after serious hepatotoxicity developed.

Topics: Agaricales; Amanitins; China; Drainage; Humans; Male; Middle Aged; Mushroom Poisoning; Silymarin

Topics: Amanitins; Critical Care; Humans; Liver; Mushroom Poisoning

Acute liver failure (ALF) induced by amatoxin-containing mushrooms accounts for more than 90% of deaths in patients suffering from mushroom poisoning. However, due to the fact that most hospitals cannot identify the species of mushrooms involved, or detect amatoxins, the early diagnosis of amatoxin intoxication remains a significant challenge in clinical practice.. Two patients were had ingested wild mushrooms 15 hours before admission. Six hours prior to admission they experienced nausea, vomiting, weakness, abdominal pain and diarrhea. The species of mushrooms they had consumed could not be identified.. According to their delayed gastroenteritis, the two patients were clinically diagnosed with amatoxin poisoning. One week after the patients were discharged, the species of the mushrooms was identified as Amanita fuliginea and the diagnosis was confirmed.. The two patients were treated with silibinin, penicillin G and plasma exchange.. Although the two patients progressed to ALF they fully recovered and were discharged on day 10 after admission.. Our case reports suggested that patients with unidentified wild mushroom intoxication with delayed gastroenteritis could be clinically diagnosed with amatoxin poisoning; in such cases, liver coagulation function should be frequently evaluated. Early diagnosis and treatment are crucial for survival in patients with ALF induced by amatoxin poisoning.

Topics: Adult; Amanita; Amanitins; Anti-Bacterial Agents; Antioxidants; Diagnosis, Differential; Early Diagnosis; Female; Humans; Liver Failure, Acute; Middle Aged; Mushroom Poisoning; Penicillin G; Plasma Exchange; Silybin; Silymarin

Acute liver failure (ALF) is associated with high mortality, and a poor understanding of the underlying pathophysiology has resulted in a lack of effective treatments so far. Here, using an amatoxin-induced rhesus monkey model of ALF, we panoramically revealed the cellular and molecular events that lead to the development of ALF. The challenged monkeys with toxins underwent a typical course of ALF including severe hepatic injury, systemic inflammation and eventual death. Adaptive immune was not noticeably disturbed throughout the progress of ALF. A systematic examination of serum factors and cytokines revealed that IL-6 increase was the most rapid and drastic. Interestingly, we found that IL-6 was mainly produced by circulating monocytes. Furthermore, ablation of monocyte-derived IL-6 in mice decreased liver injury and systemic inflammation following chemical injection. Our findings reveal a critical role of circulating monocytes in initiating and accelerating ALF, indicating a potential therapeutic target in clinical treatment for ALF.

Topics: Alanine Transaminase; Amanitins; Animals; Aspartate Aminotransferases; Cytokines; Disease Progression; Gene Expression; Hepatic Encephalopathy; Interleukin-6; L-Lactate Dehydrogenase; Lipopolysaccharides; Liver Failure, Acute; Liver Function Tests; Macaca mulatta; Mice; Monocytes

Cases of mushroom poisoning in Thailand have increased annually. During 2008 to 2014, the cases reported to the National Institute of Health included 57 deaths; at least 15 died after ingestion of amanitas, the most common lethal wild mushrooms inhabited. Hence, the aims of this study were to identify mushroom samples from nine clinically reported cases during the 7-year study period based on nuclear ITS sequence data and diagnose lethal peptide toxins using a reversed phase LC-MS method. Nucleotide similarity was identified using BLAST search of the NCBI database and the Barcode of Life Database (BOLD). Clade characterization was performed by maximum likelihood and Bayesian phylogenetic approaches. Based on BLAST and BOLD reference databases our results yielded high nucleotide similarities of poisonous mushroom samples to A. exitialis and A. fuliginea. Detailed phylogenetic analyses showed that all mushroom samples fall into their current classification. Detection of the peptide toxins revealed the presence of amatoxins and phallotoxins in A. exitialis and A. fuliginea. In addition, toxic α-amanitin was identified in a new provisional species, Amanita sp.1, with the highest toxin quantity. Molecular identification confirmed that the mushrooms ingested by the patients were members of the lethal amanitas in the sections Amanita and Phalloideae. In Thailand, the presence of A. exitialis was reported here for the first time and all three poisonous mushroom species provided new and informative data for clinical studies.

Topics: Amanita; Amanitins; Chromatography, Liquid; Databases, Genetic; Humans; Mass Spectrometry; Mushroom Poisoning; Retrospective Studies; Sequence Analysis, DNA; Thailand

Published estimates of survival associated with mushroom (amatoxin)-induced acute liver failure (ALF) and injury (ALI) with and without liver transplant (LT) are highly variable. We aimed to determine the 21-day survival associated with amatoxin-induced ALI (A-ALI) and ALF (A-ALF) and review use of targeted therapies.. Cohort study of all A-ALI/A-ALF patients enrolled in the US ALFSG registry between 01/1998 and 12/2014.. Of the 2224 subjects in the registry, 18 (0.8%) had A-ALF (n = 13) or A-ALI (n = 5). At admission, ALF patients had higher lactate levels (5.2 vs. 2.2 mm, P = 0.06) compared to ALI patients, but INR (2.8 vs. 2.2), bilirubin (87 vs. 26 μm) and MELD scores (28 vs. 24) were similar (P > 0.2 for all). Of the 13 patients with ALF, six survived without LT (46%), five survived with LT (39%) and two died without LT (15%). Of the five patients with ALI, four (80%) recovered and one (20%) survived post-LT. Comparing those who died/received LT (non-spontaneous survivors [NSS]) with spontaneous survivors (SS), N-acetylcysteine was used in nearly all patients (NSS 88% vs. SS 80%); whereas, silibinin (25% vs. 50%), penicillin (50% vs. 25%) and nasobiliary drainage (0 vs. 10%) were used less frequently (P > 0.15 for all therapies).. Patients with mushroom poisoning with ALI have favourable survival, while around half of those presenting with ALF may eventually require LT. Further study is needed to define optimal management (including the use of targeted therapies) to improve survival, particularly in the absence of LT.

Topics: Acetylcysteine; Adult; Amanitins; Chemical and Drug Induced Liver Injury; Cohort Studies; Female; Humans; Liver Failure, Acute; Liver Transplantation; Male; Middle Aged; Mushroom Poisoning; North America; Penicillins; Registries; Silybin; Silymarin

Mushroom poisoning is a cause of major mortality and morbidity all over the world. Although Hong Kong people consume a lot of mushrooms, there are only a few clinical studies and reviews of local mushroom poisoning. This study aimed to review the clinical characteristics, source, and outcome of mushroom poisoning incidences in Hong Kong.. This descriptive case series review was conducted by the Hong Kong Poison Information Centre and involved all cases of mushroom poisoning reported to the Centre from 1 July 2005 to 30 June 2015.. Overall, 67 cases of mushroom poisoning were reported. Of these, 60 (90%) cases presented with gastrointestinal symptoms of vomiting, diarrhoea, and abdominal pain. Gastrointestinal symptoms were early onset (<6 hours post-ingestion) and not severe in 53 patients and all recovered after symptomatic treatment and a short duration of hospital care. Gastrointestinal symptoms, however, were of late onset (≥6 hours post-ingestion) in seven patients; these were life-threatening cases of amatoxin poisoning. In all cases, the poisonous mushroom had been picked from the wild. Three cases were imported from other countries, and four collected and consumed the amatoxin-containing mushrooms in Hong Kong. Of the seven cases of amatoxin poisoning, six were critically ill, of whom one died and two required liver transplantation. There was one confirmed case of hallucinogenic mushroom poisoning caused by Tylopilus nigerrimus after consumption of a commercial mushroom product. A number of poisoning incidences involved the consumption of wild-harvested dried porcini purchased in the market.. Most cases of mushroom poisoning in Hong Kong presented with gastrointestinal symptoms and followed a benign course. Life-threatening cases of amatoxin poisoning are occasionally seen. Doctors should consider this diagnosis in patients who present with gastrointestinal symptoms that begin 6 hours or more after mushroom consumption.

Topics: Abdominal Pain; Adolescent; Adult; Aged; Aged, 80 and over; Amanitins; Child; Child, Preschool; Diarrhea; Female; Gastrointestinal Diseases; Hong Kong; Humans; Male; Middle Aged; Mushroom Poisoning; Retrospective Studies; Vomiting; Young Adult

BACKGROUND Fractionated plasma separation and absorption (FPSA) is an extracorporeal liver support method that detoxifies accumulated toxins. There are limited data of its use in the treatment of Amanita phalloides intoxication. The objective of this study was to investigate whether FPSA before liver transplantation improves patients' short-term post liver transplantation survival in Amanita phalloides poisoning. MATERIAL AND METHODS The study population consisted of ten patients who had liver transplantation (LT) due to acute liver failure (ALF) caused by Amanita phalloides poisoning. Six patients were treated with FPSA before liver transplantation. All the patients who were started on FPSA were also placed on the liver transplantation list according to emergent liver transplantation criteria. RESULTS Patients treated with FPSA were in a more severe clinical condition presenting in higher mean MELD, total bilirubin, INR and ammonia along with more frequent hypoglycemia and hepatic encephalopathy grade 3/4. FPSA group had longer mean waiting time on the recipient list (3.5 vs. 1.25 days) but inferior thirty-day survival rate (16.5% vs. 100%). CONCLUSIONS When conservative medical modalities are ineffective, the only treatment for Amanita phalloides poisoning is a liver transplant. Although FPSA treated patients had inferior post-LT survival, FPSA was found to prolong the pre surgical waiting time for critically ill patients, consequently giving a chance of life-saving procedure.

Topics: Adult; Aged; Amanita; Amanitins; Female; Humans; Kaplan-Meier Estimate; Liver Failure, Acute; Liver Transplantation; Male; Middle Aged; Mushroom Poisoning; Retrospective Studies; Sorption Detoxification; Time Factors; Waiting Lists; Young Adult

Lethal amanitas (Amanita sect. Phalloideae) are responsible for 90% of all fatal mushroom poisonings. Since 2000, more than ten new lethal Amanita species have been discovered and some of them had caused severe mushroom poisonings in China. However, the contents and distribution of cyclopeptides in these lethal mushrooms remain poorly known. In this study, the diversity of major cyclopeptide toxins in seven Amanita species from Eastern Asia and three species from Europe and North America were systematically analyzed, and a new approach to inferring phylogenetic relationships using cyclopeptide profile was evaluated for the first time. The results showed that there were diversities of the cyclopeptides among lethal Amanita species, and cyclopeptides from Amanita rimosa and Amanita fuligineoides were reported for the first time. The amounts of amatoxins in East Asian Amanita species were significantly higher than those in European and North American species. The analysis of distribution of amatoxins and phallotoxins in various Amanita species demonstrated that the content of phallotoxins was higher than that of amatoxins in Amanita phalloides and Amanita virosa. In contrast, the content of phallotoxins was significantly lower than that of amatoxins in all East Asian lethal Amanita species tested. However, the distribution of amatoxins and phallotoxins in different tissues showed the same tendency. Eight cyclopeptides and three unknown compounds were identified using cyclopeptide standards and high-resolution MS. Based on the cyclopeptide profiles, phylogenetic relationships of lethal amanitas were inferred through a dendrogram generated by UPGMA method. The results showed high similarity to the phylogeny established previously based on the multi-locus DNA sequences.

Topics: Amanita; Amanitins; Chromatography, High Pressure Liquid; Mass Spectrometry; Mushroom Poisoning; Peptides, Cyclic; Phylogeny; Reference Standards

Most of the fatal cases of mushroom poisoning are caused by Amanita phalloides. The amount of toxin in mushroom varies according to climate and environmental conditions. The aim of this study is to measure α-, β-, and γ-amanitin with phalloidin and phallacidin toxin concentrations. Six pieces of A. phalloides mushrooms were gathered from a wooded area of Düzce, Turkey, on November 23, 2011. The mushrooms were broken into pieces as spores, mycelium, pileus, gills, stipe, and volva. α-, β-, and γ-Amanitin with phalloidin and phallacidin were analyzed using reversed-phase high-performance liquid chromatography. As a mobile phase, 50 mM ammonium acetate + acetonitrile (90 + 10, v/v) was used with a flow rate of 1 mL/min. C18 reverse phase column (150 × 4.6 mm; 5 µm particle) was used. The least amount of γ-amanitin toxins was found at the mycelium. The other toxins found to be in the least amount turned out to be the ones at the spores. The maximum amounts of amatoxins and phallotoxin were found at gills and pileus, respectively. In this study, the amount of toxin in the spores of A. phalloides was published for the first time, and this study is pioneering to deal with the amount of toxin in mushrooms grown in Turkey.

Topics: Alpha-Amanitin; Amanita; Amanitins; Chromatography, High Pressure Liquid; Chromatography, Reverse-Phase; Forests; Fruiting Bodies, Fungal; Humans; Mushroom Poisoning; Mycelium; Peptides, Cyclic; Phalloidine; Species Specificity; Spectrophotometry, Ultraviolet; Spores, Fungal; Turkey

Serious to fatal toxicity may occur with amanitin-containing mushrooms ingestions. A Lepiota brunneoincarnata familial poisoning with hepatic toxicity is reported. In such poisonings, acute gastroenteritis may be firstly misdiagnosed leading to delay in preventing liver dysfunction by silibinin or penicillin G. Mushroom picking finally requires experience and caution.

Topics: Administration, Oral; Adult; Agaricales; Amanitins; Antidotes; Charcoal; Child; Diagnosis, Differential; Emergency Service, Hospital; Female; Fluid Therapy; Follow-Up Studies; Humans; Male; Mushroom Poisoning

Mushroom poisonings occur when ingestion of wild mushrooms containing toxins takes place, placing the consumers at life-threatening risk. In the present case report, an unusual multiple poisoning with isoxazoles- and amatoxins-containing mushrooms in a context of altered mental state and poorly controlled hypertension is presented. A 68-year-old female was presented to São João hospital (Portugal) with complaints of extreme dizziness, hallucinations, vertigo and imbalance, 3 h after consuming a stew of wild mushrooms. The first observations revealed altered mental state and elevated blood pressure. The examination of cooked mushroom fragments allowed a preliminary identification of Amanita pantherina. Gas chromatography-mass spectrometry (GC-MS) showed the presence of muscimol in urine. Moreover, through high-performance liquid chromatography-ultraviolet detection (HPLC-UV) analysis of the gastric juice, the presence of α-amanitin was found, showing that amatoxins-containing mushrooms were also included in the stew. After 4 days of supportive treatment, activated charcoal, silybin and N-acetylcysteine, the patient recovered being discharged 10 days post-ingestion with no organ complications. The prompt and appropriate therapy protocol for life-threatening amatoxins toxicity probably saved the patient's life as oral absorption was decreased and also supportive care was immediately started.

Topics: Acetylcysteine; Agaricales; Aged; Alpha-Amanitin; Amanita; Amanitins; Charcoal; Female; Gas Chromatography-Mass Spectrometry; Humans; Isoxazoles; Mushroom Poisoning; Silybin; Silymarin

There are few data estimating the human lethal dose of amatoxins or of the toxin level present in ingested raw poisonous mushrooms. Here, we present a patient who intentionally ingested several wild collected mushrooms to assess whether they were poisonous. Nearly 1 day after ingestion, during which the patient had nausea and vomiting, he presented at the emergency department. His transaminase levels started to increase starting from hour 48 and peaking at hour 72 (alanine aminotransferase 2496 IU/L; aspartate aminotransferase 1777 IU/L). A toxin analysis was carried out on the mushrooms that the patient said he had ingested. With reversed-phase high-performance liquid chromatography analysis, an uptake of approximately 21.3 mg amatoxin from nearly 50 g mushroom was calculated; it consisted of 11.9 mg alpha amanitin, 8.4 mg beta amanitin, and 1 mg gamma amanitin. In the urine sample taken on day 4, 2.7 ng/mL alpha amanitin and 1.25 ng/mL beta amanitin were found, and there was no gamma amanitin. Our findings suggest that the patient ingested approximately 0.32 mg/kg amatoxin, and fortunately recovered after serious hepatotoxicity developed.

Topics: Amanita; Amanitins; Chromatography, High Pressure Liquid; Humans; Male; Middle Aged; Mushroom Poisoning

Species in the mushroom genus Lepiota can cause fatal mushroom poisonings due to their content of amatoxins such as α-amanitin. Previous studies of the toxin composition of poisonous Lepiota species relied on analytical methods of low sensitivity or resolution. Using liquid chromatography coupled to UV absorbance and mass spectrometry, we analyzed the spectrum of peptide toxins present in six Italian species of Lepiota, including multiple samples of three of them collected in different locations. Field taxonomic identifications were confirmed by sequencing of the internal transcribed spacer (ITS) regions. For comparison, we also analyzed specimens of Amanita phalloides from Italy and California, a specimen of A. virosa from Italy, and a laboratory-grown sample of Galerina marginata. α-Amanitin, β-amanitin, amanin, and amaninamide were detected in all samples of L. brunneoincarnata, and α-amanitin and γ-amanitin were detected in all samples of L. josserandii. Phallotoxins were not detected in either species. No amatoxins or phallotoxins were detected in L. clypeolaria, L. cristata, L. echinacea, or L. magnispora. The Italian and California isolates of A. phalloides had similar profiles of amatoxins and phallotoxins, although the California isolate contained more β-amanitin relative to α-amanitin. Amaninamide was detected only in A. virosa.

Topics: Agaricales; Amanitins; Base Sequence; Chromatography, High Pressure Liquid; DNA, Fungal; Mass Spectrometry; Molecular Sequence Data; Sequence Analysis, DNA; Spectrophotometry, Ultraviolet

Amatoxins are ribosomally encoded and posttranslationally modified peptides that account for the majority of fatal mushroom poisonings of humans. A representative amatoxin is the bicyclic octapeptide α-amanitin, formed via head-to-tail macrocyclization, which is ribosomally biosynthesized as a 35-amino acid propeptide in Amanita bisporigera and in the distantly related mushroom Galerina marginata. Although members of the prolyl oligopeptidase (POP) family of serine proteases have been proposed to play a role in α-amanitin posttranslational processing, the exact mechanistic details are not known. Here, we show that a specific POP (GmPOPB) is required for toxin maturation in G. marginata. Recombinant GmPOPB catalyzed two nonprocessive reactions: hydrolysis at an internal Pro to release the C-terminal 25-mer from the 35-mer propeptide and transpeptidation at the second Pro to produce the cyclic octamer. On the other hand, we show that GmPOPA, the putative housekeeping POP of G. marginata, behaves like a conventional POP.

Topics: Alpha-Amanitin; Amanita; Amanitins; Amino Acid Sequence; Biocatalysis; Cyclization; Kinetics; Molecular Sequence Data; Mutation; Prolyl Oligopeptidases; Serine Endopeptidases; Species Specificity

Topics: Amanita; Amanitins; Fatal Outcome; Female; Humans; Middle Aged; Mushroom Poisoning

Picking wild mushrooms is a popular pastime in Switzerland. Correct identification of the species is difficult for laypersons. Ingestion of toxic mushrooms may result in serious toxicity, including death. The aim of the study is to analyze and describe the circumstances of exposure to mushrooms, and to define the clinical relevance of mushroom poisoning for humans in Central Europe.. We performed a retrospective case study and analyzed all inquiries concerning human exposures to mushrooms (n = 5638, 1.2% of all inquiries) which were reported to the Swiss Toxicological Information Centre between January 1995 and December 2009.. The most frequent reason for contacting the poison center in cases of adult exposure was toxicity resulting from edible species. Pediatric exposure predominantly occurred from mushrooms found around the home. Severe symptoms have not only been observed after ingestion of non-amatoxin-containing toxic mushrooms, i.e. Boletus sp. and Cortinarius sp., but also after meals of edible species. The mortality of confirmed amatoxin poisonings was high (5/32) compared to other reports.. Inquiries regarding mushroom poisoning were a relatively infrequent reason for contacting the poison center. Nevertheless, accidental ingestion of toxic mushrooms can be responsible for severe or fatal poisonings. Although pediatric exposure to mushrooms found around the home has not led to serious toxicity in this study, prevention of exposure is warranted. Inspection of wild mushrooms by a certified mushroom expert or a mycologist seems to be a safe procedure which should be recommended.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Amanitins; Child; Child, Preschool; Eating; Female; Humans; Infant; Male; Middle Aged; Mushroom Poisoning; Poison Control Centers; Seasons; Switzerland; Young Adult

Amatoxin poisoning induces a delayed onset of acute liver failure which might be explained by the prolonged persistence of the toxin in the enterohepatic circulation. Aim of the study was to demonstrate amanitin kinetics in the enterohepatic circulation.. Four pigs underwent α-amanitin intoxication receiving 0.35 mg/kg (n=2) or 0.15 mg/kg (n=2) intraportally. All pigs remained under general anesthesia throughout the observation period of 72 h. Laboratory values and amanitin concentration in systemic and portal plasma, bile and urine samples were measured.. Amanitin concentrations measured 5h after intoxication of 219±5ng/mL (0.35 mg/kg) and 64±3 (0.15 mg/kg) in systemic plasma and 201±8ng/mL, 80±13ng/mL in portal plasma declined to baseline levels within 24h. Bile concentrations simultaneously recorded showed 153±28ng/mL and 99±58ng/mL and decreased slightly delayed to baseline within 32 h. No difference between portal and systemic amanitin concentration was detected after 24h.. Amanitin disappeared almost completely from systemic and enterohepatic circulation within 24 h. Systemic detoxification and/or interrupting the enterohepatic circulation at a later date might be poorly effective.

Topics: Alpha-Amanitin; Amanitins; Animals; Aspartate Aminotransferases; Disease Models, Animal; Enterohepatic Circulation; Female; Histocytochemistry; Liver Failure, Acute; Prothrombin Time; Swine

OBJECTIVE. We aimed to determine clinical and laboratory findings that were different between those patients who died and those who survived and to look for factors associated with the mortality in amatoxin-containing mushroom poisoning. METHODS. The mushroom poisoning patients who were admitted to our clinic between 1996 and 2009 were retrospectively evaluated. The diagnosis was based on a history of mushroom ingestion, clinical picture and the presence of serum alpha-amanitin. Patients were divided into two groups as the survival group and the fatality group. Clinical and laboratory findings were compared between the two groups. Relation between variables and clinical outcome was analyzed. RESULTS. A total of 144 amatoxin poisoning patients were included in this study. Patients who died were more likely to have demonstrated low mean arterial pressure, encephalopathy, mucosal hemorrhage, oliguria-anuria, hypoglycemia, and thrombocytopenia during the hospitalization. Low sodium values and high urea, AST, ALT, total bilirubin, LDH, prothrombin time, international normalized ratio, and activated partial thromboplastin time values were associated with increased likelihood of mortality. Nineteen patients developed acute renal failure. Fourteen patients developed acute hepatic failure. All the 14 patients who died developed acute hepatic failure. The mortality rate was 9.7%. CONCLUSIONS. The factors associated with mortality determined in this retrospective study may be helpful for clinical outcome assessment and monitoring of patients with amatoxin-containing mushroom poisoning.

Topics: Adult; Amanitins; Female; Hemoperfusion; Humans; Liver Failure, Acute; Male; Middle Aged; Mushroom Poisoning; Retrospective Studies

Some species in the genus Amanita have a great variety of toxic secondary metabolites. They are characterized macroscopically by having a white spore print and free gills, and microscopically by the presence of a divergent hymenophoral trama. Some species of Amanita present in Colombia were chemically characterized by analyzing their toxin composition using HPLC. Samples were collected in oak (Quercus humboldtii) and pine (Pinus radiata) forests. Twelve species were recovered, Amanita fuligineodisca, Amanita xylinivolva, Amanita flavoconia, Amanita rubescens, Amanita bisporigera, Amanita muscaria, Amanita humboldtii, Amanita sororcula, Amanita brunneolocularis, Amanita colombiana, Amanita citrina, Amanita porphyria as well as two unreported species. Results showed that most of the analyzed species have α -amanitin in concentrations ranging from 50 ppm to 6000 ppm. Concentrations of α-amanitin in the pileus were significantly greater than in the stipe. Phalloidin and phallacidin were only present in A. bisporigera. Chromatographic profiles are proposed as an additional taxonomic tool since specific peaks with similar retention times were conserved at the species level.

Topics: Alpha-Amanitin; Amanita; Amanitins; Chromatography, High Pressure Liquid; Colombia; Pinus; Quercus

Ninety percent of fatal higher fungus poisoning is due to amatoxin-containing mushroom species. In addition to absence of antidote, no chemotherapeutic consensus was reported. The aim of the present study is to perform a retrospective multidimensional multivariate statistic analysis of 2110 amatoxin poisoning clinical cases, in order to optimize therapeutic decision-making. Our results allowed to classify drugs as a function of their influence on one major parameter: patient survival. Active principles were classified as first intention, second intention, adjuvant or controversial pharmaco-therapeutic clinical intervention. We conclude that (1) retrospective multidimensional multivariate statistic analysis of complex clinical dataset might help future therapeutic decision-making and (2) drugs such as silybin, N-acetylcystein and putatively ceftazidime are clearly associated, in amatoxin poisoning context, with higher level of patient survival.

Topics: Acetylcysteine; Algorithms; Amanitins; Anti-Bacterial Agents; Antioxidants; Ceftazidime; Databases, Factual; Decision Making; Decision Theory; Factor Analysis, Statistical; Humans; Multivariate Analysis; Mushroom Poisoning; Retrospective Studies; Silybin; Silymarin; Survival; Survival Rate

There is a paucity of knowledge about prenatal and perinatal risks through maternal amatoxin poisoning. No symptoms of amatoxin intoxication, except for a slight temporary increase in liver enzymes activity, occurred in a term newborn after delivery despite an Amanita phalloides intoxication of the mother 2 days before. Considering previous reports, severe fetal intoxication may not occur during the entire pregnancy.

Topics: Adult; Amanitins; Female; Humans; Infant, Newborn; Liver Function Tests; Male; Mushroom Poisoning; Pregnancy; Pregnancy Complications; Pregnancy Outcome; Risk Factors

Topics: Alpha-Amanitin; Amanitins; Animals; Antidotes; Chemical and Drug Induced Liver Injury; Cimetidine; Humans; Mice; Mushroom Poisoning

Current treatment of amatoxin poisoning includes the administration of silibinin and penicillin in combination or silibinin alone. The aim of this study was to compare both therapeutic regimes.. Of 604 patients with the suspected diagnosis of amatoxin poisoning 367 were retrospectively analysed: 118 patients had received silibinin alone and 249 patients silibinin in combination with penicillin. Logistic regression analyses were applied to investigate the efficacy of both therapeutic regimens by comparing death and liver transplantation rates. A potentially independent effect on outcome of age, sex, year of treatment, latency period of symptoms and start of silibinin therapy was taken into account.. In the group who had received the combination of silibinin and penicillin 8.8% died or underwent liver transplantation compared to 5.1% in the group of those who had received silibinin alone. The risk of death or organ transplantation was thus reduced by nearly 40% in the latter group (adjusted odds ratio: 0.58; 95% CI: 0.21-1.57; p=0.28). A longer latency period (< or =12h vs. >12h) was associated with a significant reduction of this risk (adjusted OR.: 6.10; 95% CI:1.77-21.3; p=0.004). A later start of silibinin therapy (>24h vs. < or = 24h) was associated with a tendency toward an increased frequency of death or organ transplantation (adjusted OR.: 3.0; 95% CI: 0.96-9.20; p=0.059).. A lower death and transplantation rate was observed in the silibinin treatment group than in group treated with silibinin combined with penicillin. However, this difference was not statistically significant. The high risk ratio relating to the time-dependent effect of silibinin suggests its efficaciousness in the treatment of amatoxin poisoning. The latency period was assessed as an independent prognostic factor.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Amanita; Amanitins; Antidotes; Antioxidants; Child; Child, Preschool; Drug Therapy, Combination; Female; Humans; Liver Transplantation; Logistic Models; Male; Middle Aged; Mushroom Poisoning; Penicillins; Prognosis; Retrospective Studies; Severity of Illness Index; Silybin; Silymarin; Time Factors

Two patients, a 54-year-old man and a 51-year-old woman, presented with abdominal pain, vomiting and diarrhoea; these symptoms developed 9 and 15 hours, respectively, after consumption of soup from mushrooms that they had picked themselves. As a result of these events, a third patient, a 55-year-old woman with diarrhoea who had also eaten the soup, also presented herself. The first patient recognised deathcap or death angel mushrooms (Amanita phalloides) on a photograph. All three patients were treated for amatoxin poisoning with a combination of high-dose penicillin G, silibinin and acetylcysteine intravenously. The poisoning was later confirmed by the results of urinalysis. The patients were discharged in good condition 8 days later.

Topics: Acetylcysteine; Amanita; Amanitins; Antioxidants; Drug Therapy, Combination; Female; Humans; Male; Middle Aged; Mushroom Poisoning; Penicillin G; Silybin; Silymarin; Treatment Outcome; Urinalysis

The foraging of wild mushrooms can be complicated by toxicity from several mushroom types. Amatoxin, a peptide contained in several mushroom species, accounts for the majority of severe mushroom poisonings by binding to RNA polymerase II irreversibly, leading to severe hepatonecrosis. There is no effective antidote for severe amatoxin poisoning. We compare the effectiveness of 5 potential antidotal therapies in limiting the degree of hepatonecrosis in a randomized, controlled, murine model of amatoxin-induced hepatotoxicity.. One hundred eighty male Institute of Cancer Research mice were randomized into 6 equal groups. Within each group, 21 mice were intraperitoneally injected with 0.6 mg/kg of alpha-amanitin (amatoxin); the remaining 9 were injected with 0.9% normal saline solution. Four hours postinjection, each group of 30 mice was randomized to 1 of 5 intraperitoneal treatments (N-acetylcysteine, benzylpenicillin, cimetidine, thioctic acid, or silybin) or normal saline solution. Repeated dosing was administered intraperitoneally every 4 to 6 hours for 48 hours. After 48 hours of treatment, each subject was killed, cardiac blood was aspirated for hepatic aminotransferase measurements (alanine transaminase and aspartate transaminase), and liver specimens were harvested to evaluate the extent of hepatonecrosis. The degree of hepatonecrosis was determined by a pathologist blinded to the treatment group and divided into 5 categories according to percentage of hepatonecrosis.. Amanitin significantly increased aspartate transaminase in treated mice compared with normal saline solution-treated controls (mean [SD] 2,441 [2,818] IU/L versus 310 [252]; P=.03). None of the antidotal therapies were found to significantly decrease the increase in aminotransferases compared with controls. Further, none of the antidotal therapies demonstrated an important decrease in hepatonecrosis compared with controls when a histologic grading scale was used.. In this murine model, N-acetylcysteine, benzylpenicillin, cimetidine, thioctic acid, and silybin were not effective in limiting hepatic injury after alpha-amanitin poisoning. Increases of aminotransferases and degrees of histologic hepatonecrosis were not attenuated by these antidotal therapies.

Topics: Acetylcysteine; Alanine Transaminase; Amanitins; Animals; Antidotes; Aspartate Aminotransferases; Chemical and Drug Induced Liver Injury; Cimetidine; Disease Models, Animal; Male; Mice; Mushroom Poisoning; Penicillin G; Random Allocation; Silybin; Silymarin; Thioctic Acid

Topics: Amanitins; Creatinine; Emergency Medical Services; Humans; Liver Transplantation; Mushroom Poisoning; Predictive Value of Tests; Prothrombin

Amatoxins, the lethal constituents of poisonous mushrooms in the genus Amanita, are bicyclic octapeptides. Two genes in A. bisporigera, AMA1 and PHA1, directly encode alpha-amanitin, an amatoxin, and the related bicyclic heptapeptide phallacidin, a phallotoxin, indicating that these compounds are synthesized on ribosomes and not by nonribosomal peptide synthetases. alpha-Amanitin and phallacidin are synthesized as proproteins of 35 and 34 amino acids, respectively, from which they are predicted to be cleaved by a prolyl oligopeptidase. AMA1 and PHA1 are present in other toxic species of Amanita section Phalloidae but are absent from nontoxic species in other sections. The genomes of A. bisporigera and A. phalloides contain multiple sequences related to AMA1 and PHA1. The predicted protein products of this family of genes are characterized by a hypervariable "toxin" region capable of encoding a wide variety of peptides of 7-10 amino acids flanked by conserved sequences. Our results suggest that these fungi have a broad capacity to synthesize cyclic peptides on ribosomes.

Topics: Amanita; Amanitins; Amino Acid Sequence; Base Sequence; Basidiomycota; Conserved Sequence; Fungal Proteins; Genes, Fungal; Molecular Sequence Data; Multigene Family; Peptides, Cyclic; Prolyl Oligopeptidases; Protein Structure, Tertiary; Ribosomes; Sequence Alignment; Sequence Homology, Amino Acid; Serine Endopeptidases; Species Specificity

Topics: Amanitins; Humans; Liver Failure, Acute; Mushroom Poisoning

Indication of liver transplantation in acute liver failure following amatoxin intoxication is still uncertain.. One hundred and ninety-eight patients were studied retrospectively. The laboratory parameters alanine-aminotransferase, serum bilirubin, serum creatinine and prothrombin index were analyzed over time. Predictors of fatal outcome and survival were determined by receiver-operating-characteristic and sensitivity-specificity analysis.. Twenty-three patients died in the median 6.1 days (range, 2.7-13.9 days) after ingestion. Using a single parameter as predictor of fatal outcome the area under the receiver-operating-characteristic curve of prothrombin index (0.96) and serum creatinine (0.93) were both significantly greater (P<0.05) compared with serum bilirubin (0.82) and alanine-aminotransferase (0.69). Prediction of fatal outcome had an optimum, if a prothrombin index less than 25% was combined with a serum creatinine greater than 106 micromol/l from day 3 after ingestion onwards (sensitivity 100%, 95% confidence interval 87-100; specificity 98%, 95% confidence interval 94-100). The median time period between the first occurrence of this predictor in non-survivors and death was 63h (range, 3-230h).. A decision model of liver transplantation following amatoxin intoxication using prothrombin index in combination with serum creatinine from day 3 to 10 after ingestion enables an early and reliable assessment of outcome.

Topics: Adolescent; Adult; Alanine Transaminase; Amanitins; Bilirubin; Child; Humans; Liver Failure; Liver Transplantation; Mushroom Poisoning; Predictive Value of Tests; Prothrombin Time; Retrospective Studies; Survival Analysis

Topics: Amanitins; Humans; Liver Transplantation; Risk; Unnecessary Procedures

Amanita exitialis Zhu L. Yang and T.H. Li is a lethal mushroom species recently isolated in Guangdong Province, China. In this report, a pure culture of this species was obtained for the first time. To confirm the identity of the pure culture, the internal transcribed spacer regions of the nuclear ribosomal DNA of the pure culture and of a typical fruiting body of the species were sequenced and compared. Further, amatoxins produced by pure cultures were analyzed and characterized by high-performance liquid chromatography and mass spectrometry analysis. The results showed that the pure cultures produced 728.3 +/- 43.8 microg g(-1) (dry matter) of alpha-Amanitin and 60.0 +/- 20.7 microg g(-1) (dry matter) of beta-Amanitin, respectively, a yield which is about 10% of that produced by fruiting bodies.

Topics: Amanita; Amanitins; Chromatography, High Pressure Liquid; DNA, Fungal; DNA, Intergenic; Mass Spectrometry; Molecular Sequence Data; Mycotoxins; Photomicrography; Phylogeny; Sequence Analysis, DNA

This paper presents species of fungi of high toxicity. Their consumption might have serious consequences for health and in many cases it might lead to death. Toxic compounds present in fungi have also been characterised, mechanisms of their toxic activity have been presented and clinical symptoms of poisoning have been described. Hallucinogenic mushrooms have also been mentioned as they have recently become a serious problem: many people use them to intoxicate themselves. There are also species of mushrooms that can be consumed under certain conditions since they can occasionally trigger off serious disturbances for the functioning of organisms.

Topics: Amanita; Amanitins; Cross-Sectional Studies; Humans; Incidence; Mushroom Poisoning; Peptides, Cyclic; Poland; Structure-Activity Relationship

Experiences of liver transplantation after Amanita phalloides poisoning were analysed in anaesthetic and intensive therapist point of view based on 3 cases. Cardiac problems were found at all patients during the postoperative period. Probably the amatoxin has cardiotoxic effect or a part of phallotoxins are absorbed despite cooking and caused reversible cardiac function impairment. Pancreatitis, DIC, gastrointestinal bleeding, acute renal failure were found at all patients, therefore liver transplantation is only a part of the treatment, complex therapy is necessary in this cases.

Topics: Acute Kidney Injury; Adult; Amanita; Amanitins; Anesthesia, General; Child; Combined Modality Therapy; Critical Care; Disseminated Intravascular Coagulation; Electrocardiography; Gastrointestinal Hemorrhage; Heart Conduction System; Humans; Liver Failure; Liver Transplantation; Male; Mushroom Poisoning; Pancreatitis

Conocybe lactea was examined as part of a larger study on the distribution of amatoxins and phallotoxins in fungi, and the taxonomic relationships between these fungi. As amatoxins are present in the congener C. filaris, the locally abundant C. lactea was examined using HPLC and mass spectroscopy. Amatoxins were not found in C. lactea, but the related phallotoxins were present in small quantities making it the first fungus outside of the genus Amanita in which phallotoxins have been detected. Despite the presence of a related toxin, C. lactea was found not to be taxonomically close to C. filaris. Phylogenetic analyses using nuclear ribosomal RNA genes indicated that North American specimens of C. lactea were conspecific with North American specimens of C. crispa in Conocybe sect. Candidae. European C. crispa was a distinct taxon. The implications of the use of the name C. albipes for these taxa are discussed. Nucleotide data confirmed placement of the sequestrate taxon Gastrocybe lateritia in sect. Candidae, but as a distinct taxon. It is hypothesized that the unique sequestrate morphology of G. lateritia may be caused by a bacterial infection.

Topics: Agaricales; Amanitins; Chromatography, High Pressure Liquid; DNA, Fungal; DNA, Ribosomal Spacer; Genes, rRNA; Mass Spectrometry; Molecular Sequence Data; Phylogeny; RNA, Ribosomal, 28S; RNA, Ribosomal, 5.8S; Sequence Analysis, DNA

The content and distribution of the main Amatoxins (alpha-amanitin, beta-amanitin) and Phallotoxins (Phallacidin, Phallisin, Phalloin, Phalloidin) in the three tissues (cap, stipe and volva) of Amanita exitialis were evaluated by means of high-performance liquid chromatography. The results showed that Amanita exitialis was a lethal mushrooms, the cap had the highest content of total toxins, it reached 8152.6 microg/g dry weight, the toxins content in stipe reached 3742.3 microg/g dry weight, whereas the volva had the lowest content of total toxins,it had only 1142.5 microg/g dry weight. The distribution of Amatoxins and Phallotoxins in the tissues were revealed and it displayed that the content of Amatoxins (alpha-amanitin and beta-amanitins especially alpha-amanitin) in the cap, stipe or volva of A. exitialis was higher than that of Phallotoxins (Phallacidin, Phallisin, Phalloidin and Phalloin). But the content of Phallotoxins especially Phallacidin was gradationally higher from cap to stipe and to volva.

Topics: Amanita; Amanitins; China; Chromatography, High Pressure Liquid; Fungal Structures

The purpose of this study was to develop a clinically relevant porcine model of fulminant hepatic failure (FHF) by means of administration of amatoxin and endotoxin.. Pigs were intraportally administered only saline in group 1 (n = 3), 1 microg/kg of lipopolysaccharide (LPS) in group 2 (n = 4), 0.1 mg/kg of alpha-amanitin in group 3 (n = 5), and amanitin plus LPS in group 4 (n = 9).. All the pigs in groups 1 and 2 survived with minimal changes in liver function tests. In contrast to the 60% mortality in group 3, all the pigs in group 4 died within 96 h, with a significant increase in aspartate transaminase at 24 h (9,757 +/- 2,167 IU/I). In addition, they demonstrated severe metabolic disorders, such as serum lactate accumulation, hypoglycemia, coagulopathy, plasma amino acid imbalance, and hyperammonemia. The intracranial pressure significantly increased to 17.8 +/- 2.5 mmHg immediately before death. Reversal of FHF in these pigs following orthotopic liver transplantation confirmed that the toxicity is liver-specific and that the graft liver is unaffected.. This porcine model of FHF induced by a combination of amanitin and LPS will be of much use in the development of new therapies for human FHF.

Topics: Amanitins; Animals; Disease Models, Animal; Humans; Intracranial Hypertension; Lipopolysaccharides; Liver; Liver Failure; Liver Transplantation; Male; Swine

We report here three cases of Amanita virosa induced toxic hepatitis. Two of the three cases recovered but the other died 10 days after mushroom ingestion. Since the mortality of Amanita mushroom induced toxic hepatitis is very high, prompt diagnosis and aggressive therapeutic measures should be initiated as soon as possible. Our cases showed that the initial serum aminotransferase levels might not predict the clinical outcome of the patient, but that the prothrombin time (PT) seemed to be a more useful prognostic marker. Close monitoring of aminotransferase levels and PT as well as appropriate therapy are recommended. All three cases showed signs of proteinuria and we were able to characterize mixed tubular and glomerular type proteinuria at 3 or 4 days after ingestion in two cases. Among the previously reported Korean cases of suspected Amanita induced toxic hepatitis, most species could not be identified except for four cases of Amanita virosa. No cases of Amanita phalloides induced toxic hepatitis have been identified in Korea so far.

Topics: Adult; Amanita; Amanitins; Chemical and Drug Induced Liver Injury; Female; Humans; Male; Middle Aged; Mushroom Poisoning; Proteinuria

Topics: Adolescent; Adult; Aged; Amanita; Amanitins; California; Cause of Death; Charcoal; Child; Colic; Diarrhea; Female; Fluid Therapy; Histamine H2 Antagonists; Humans; Male; Middle Aged; Mushroom Poisoning; Nausea; Vomiting

The toxic peptides in Amanita virosa and Amanita verna from various region were analyzed by a reversed-phase high-performance liquid chromatography. It is showed that there were great differences in the concentrations of toxic peptides between species and strains of A. virosa and A. verna.

Topics: Amanita; Amanitins; Chromatography, High Pressure Liquid; Peptides; Species Specificity

Four cases of severe Lepiota poisoning, including three which developed toxic fulminant hepatitis treated by orthotopic hepatic transplantation, are reported here. The toxicity of the Lepiota is discussed as well as the indications for hepatic transplantation in poisonings due to amatoxin-containing mushrooms.

Topics: Adult; Amanitins; Child; Electroencephalography; Female; Humans; Liver Failure; Liver Transplantation; Male; Mushroom Poisoning

We report four cases of poisoning with amatoxin-producing mushrooms in suburban Long Island. All occurred when amateur mushroom hunters picked mushrooms from neighboring lawns. Two patients presented 30 hours post ingestion with evidence of acute hepatic dysfunction. One survived, after treatment with charcoal and penicillin; the other, a 90-year-old woman with prior cardiac disease soon developed shock and subsequently died. The other two patients were admitted 18 hours after ingestion of Lepiota chlorophyllum and received prompt charcoal hemoperfusion. Both did well, although one had a mild elevation of transaminases. Although most reports of amatoxin poisoning originate in Europe, these cases confirm that amatoxin-producing mushrooms, including Lepiota chlorophyllum, may be found in northeastern American suburbs. Such patients who present prior to 24 hours after ingestion should receive charcoal hemoperfusion if a lethal dose (> 50 g of mushroom) has been eaten.

Topics: Adult; Aged; Aged, 80 and over; Amanita; Amanitins; Basidiomycota; Fatal Outcome; Female; Heart Block; Humans; Hypertension; Male; Middle Aged; Mushroom Poisoning; New York; Thyroiditis, Autoimmune

Mushroom poisonings caused by amatoxins are mostly lethal. Information about mycetisms caused by white species of Amanita is scarce. The present paper describes a case of mushroom poisoning caused by A. virosa. A prolongated latency period (6-10 hours), followed by cholera-like, improvement and visceral complication phases confirmed the amatoxin poisoning. The consumption of about 3 pounds of the toadstool by seven persons caused the death of five. Two patients survive the ingestion.

Topics: Adolescent; Adult; Aged; Amanita; Amanitins; Fatal Outcome; Female; Humans; Male; Mexico; Mushroom Poisoning

Four amatoxin-binding proteins with KD values in the nanomolar range, 3 monoclonal antibodies and RNA polymerase II, were studied with respect to their affinities to 24 alpha-amanitin derivatives with modified side chains. From KD values we estimated the amounts of binding energy that single side chains of the amatoxins contribute to complex formation. Ile6, previously identified by X-ray analysis to be part of a beta-turn (Kostansek EC, Lipscomb WN, Yocum RR, Thiessen WE, 1978, Biochemistry 17:3790-3795) proved to be of outstanding importance in all complexes. Replacement of the isoleucine with alanine reduced the affinity to all binding proteins to < 1%, suggesting a strong hydrophobic interaction. A strong effect was also seen when Gly5 was replaced with alanine, suggesting that the absence of a side chain in proximity to the beta-turn is likewise important. In addition to the beta-turn, each of the proteins showed at least 2 other points of strong contact formed by hydrogen bonds. Donors are the indole NH of 6'-hydroxy-Trp4 and OH of hydroxy-Pro2 and dihydroxy-Ile3. All the antibodies, but not RNA polymerase II, recognized the indole nucleus of 6'-hydroxy-Trp4. The geometric arrangement of the 4 strongest contact points suggests that the amatoxin binding site is different in each of the 4 proteins, except for the 2 antibodies raised in the same animal. Here, most of the contact points were identical but differed in strength of interaction. The method of structural analysis presented in this study is useful for identifying contact sites in complexes of proteins with peptides of rigid conformation. Furthermore, the method complements X-ray data by providing information on the amount of binding energy contributed by single structural elements.

Topics: Amanitins; Amino Acid Sequence; Animals; Antibodies, Monoclonal; Binding Sites; Mice; Models, Molecular; Molecular Sequence Data; Molecular Structure; Protein Binding; Protein Structure, Secondary; Rats; RNA Polymerase II; Thermodynamics

Haemoperfusion was introduced in the treatment of intoxications with Amanita phalloides in 1974. Haemoperfusion over active charcoal is used most frequently. The objective of the present study was to compare the sorption capacity of Czech haemoperfusion sorbents of the charcoal and resin type for amatoxins extracted from dried Amanita phalloides.. Haemoperfusion sorbents on the basis of charcoal-Chemviron SC XII and on the basis of synthetic resin--Amberlite XAD 2 were used in experiments in vitro. Recirculation of an aqueous extract of Amanita phalloides over a haemoperfusion column with a volume of 400 ml located in a closed system took 4 hours. Amanitin levels (alpha and beta) were assessed by HPLC. Two-hour perfusion over Amberlite XAD led to removal of the entire amount of both amanitins from the aqueous solution. Four-hour perfusion over charcoal, however, reduced the original alpha amanitin level by 24% (p < 0.05) and of beta amanitin by 8% (change not significant). Extraction of alpha and beta amanitin by the resin sorbent varied between 0.95-1.00. The type of charcoal used, produced from the shells of coconuts, had a low extraction activity, cca 12%.. The assembled results provide evidence that the synthetic resin Amberlite XAD 2 has a more than ten times greater sorption capacity, as compared with charcoal (Chemviron SC XII) and that in experiments in vitro it removes rapidly and effectively alpha and beta amanitin from an aqueous solution. Although during sorption of mycotoxins from plasma or blood the sorption rate and total capacity declines amberlite resins (Czech haemoperfusion column Hemabsorb A2) can be recommended for use in clinical toxicology.

Topics: Absorption; Amanitins; Charcoal; Hemoperfusion; In Vitro Techniques; Resins, Synthetic

Twelve patients with Galerina Autumnalis (GA) poisoning were treated. Amatoxin and phallotoxin are the principal toxins of GA. After absorption from intestine into the liver, the toxins combine with RNA polymerase, resulting in block of messenger (mRNA) synthesis, hepatocellular damage, hepatitis, hepatic necrosis, serious coagulation abnormalities and DIC. The clinical characteristics are long latent period, short period of "pseudo-remission" and serious liver dysfunction. These were pathologically confirmed by autopsy. Our experiences with this poisoning are as follows: treatment should be carried out as early as possible, especially with gastric lavage and catharsis and special attention paid to the "pseudo-remission".

Topics: Adolescent; Adult; Agaricales; Amanitins; Child; Female; Gastric Lavage; Humans; Liver; Male; Mushroom Poisoning; Necrosis

The toxin content and composition of Amanita phalloides tissues were determined in three specimens at two carpophore development stages. The carpophore was subdivided into six parts, namely, the cap, gills, ring, stipe, volva and bulb. To our knowledge, this is the first report of such an investigation on the ring and the bulb. Substantial differences in the tissue toxin content were revealed. The ring displayed a very high amount of toxins, whereas the bulb had the lowest toxin content. Compositional differences in relation to the nature of the tissue were also noted. The highest amatoxin content was found in the ring, gills and cap, whereas the bulb and volva were the richest in phallotoxins. Furthermore, variability in the toxin composition was observed. The differences in the distribution of individual toxins in the tissues might be related to the carpophore developmental stage.

Topics: Amanita; Amanitins; Chromatography, High Pressure Liquid; Hydrogen-Ion Concentration; Tissue Distribution

The kinetics of alpha and beta amanitin were studied in 45 patients intoxicated with Amanita Phalloides. The amatoxins were analyzed by high performance liquid chromatography in plasma (43 cases), urine (35 cases), gastroduodenal fluid (12 cases), feces (12 cases) and tissues (4 cases). All patients had gastrointestinal symptoms and 43 developed an acute hepatitis. Two patients underwent successful liver transplantation. Eight patients, of whom three were children, died. The detection of amatoxins in the biological fluids was time dependent. The first sample was obtained at an average of 37.9 h post ingestion in the patients with positive results and at 70.6 h in the samples without detectable amatoxins. Plasma amatoxins were detected in 11 cases at 8 to 190 ng/mL for alpha and between 23.5 to 162 ng/mL for beta. In 23 cases amatoxins were detected in urine with a mean excretion per hour of 32.18 micrograms for alpha and 80.15 micrograms for beta. In 10 patients the total amounts eliminated in the feces (time variable) ranged between 8.4 and 152 micrograms for alpha amanitin and between 4.2 and 6270 micrograms for beta amanitin. In three of four cases amatoxins were still present in the liver and the kidney after day 5. Amatoxins were usually detectable in plasma before 36 h but were present in the urine until day 4. The rapid clearance indicates that enhanced elimination of amatoxins requires early treatment. Clearance of circulating amatoxins by day 4 spares the transplanted liver.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Amanita; Amanitins; Child; Child, Preschool; Chromatography, High Pressure Liquid; Female; Humans; Liver Function Tests; Liver Transplantation; Male; Metabolic Clearance Rate; Middle Aged; Mushroom Poisoning; Prognosis; Time Factors; Tissue Distribution

The amatoxins, highly toxic components of death cap Amanita mushrooms, bind strongly to RNA polymerase II (or B) in cell nuclei thus preventing the transcription of DNAs to hn-RNAs (Pre-mRNAs), the precursors of messenger RNAs. Three of the binding sites of the bicyclic octapeptides have been identified: an isoleucine side chain in position 6, a trans-4-hydroxyl group at proline in position 2 and a hydroxylated L-isoleucine side chain in position 3. No information exists about the stereochemical conditions at the beta-C-atom (C-atom 3) of this side chain. We have now synthesized the diastereomeric S-deoxo-amaninamides (Fig. 1) containing, in position 3, L-allo-isoleucine (analog 1), (2S, 3R)-2-amino-4-hydroxy-3-methyl butyric acid (analog 2), the diastereomer (2S, 3S)-2-amino-4-hydroxy-3-methylbutyric acid (analog 3) and D-isoleucine (analog 4). In the last synthesis, besides the "normal" bicyclic octapeptide 4, an isomeric Iso-4 was formed. The affinities for Drosophila RNA polymerase II were 100 times weaker as compared to gamma-amanitin for 1, 10 times weaker for 2, 200 times weaker for 3, 100 times weaker for 4, and more than 1000 times weaker for Iso-4. The results point to the importance of a methyl group in (R)-configuration at the beta-C atom of side chain 3.

Topics: Amanitins; Amino Acid Sequence; Animals; Circular Dichroism; Drosophila melanogaster; Molecular Sequence Data; RNA Polymerase II; Structure-Activity Relationship

A reversed-phase high-performance liquid chromatographic method is described that allows the simultaneous determination of up to eight amatoxins and phallotoxins. The method identifies both neutral toxins (alpha- and gamma-amanitin, phalloidin, phallisin and phalloin) and acidic toxins (beta-amanitin, phallacidin and phallisacin). Toxins were separated, identified and determined by gradient elution with 0.02 M aqueous ammonium acetate-acetonitrile and simultaneous monitoring of the absorbances at 214 and 295 nm. The assay was successfully applied to the analysis of the toxins in a crude extract of Amanita phalloides. The limit of detection for each toxin was 10 ng/ml of extraction medium. The assay was further validated by analysing the toxin content in Galerina marginata, a species containing only amatoxins. This relatively simple method should be suitable for the detection of amatoxins and phallotoxins in almost any species of mushrooms.

Topics: Amanita; Amanitins; Chromatography, High Pressure Liquid; Quality Control

Amatoxins are cyclic peptides which can be purified from the carpophores of various mushroom species. Since they were first recognized as potent inhibitors of the nuclear RNA polymerases of most eukaryotes these peptides have served as important tools in the study of transcription. The presence of unusual amino acid residues in these peptides has provided opportunities to attempt a variety of semisynthetic modifications. We describe several new amatoxin derivatives that were prepared by selective modification of an aldehyde group which can be generated by periodate oxidation of 6'-O-methyl-alpha-amanitin. The derivatives which resulted from sodium cyanoborohydride-mediated coupling to the toxin of ammonia, glycine, and L-proline exhibited Ki values for calf thymus RNA polymerase II of 1.7 x 10(-7) M, 2.5 x 10(-7) M and 7.0 x 10(-6) M, respectively. Treatment of the aldehyde with sodium chlorite or hydroxylamine-O-sulfonic acid converted the amanitin aldehyde to the corresponding carboxyl or nitrile compounds with Ki values of 1.0 x 10(-7) M and 3.0 x 10(-9) M, respectively. Difficulties which were encountered in the preparation of these derivatives are discussed relative to peculiarities in the chemical behavior of the amanitin aldehyde.

Topics: Aldehydes; Amanitins; Circular Dichroism; Methylation; Molecular Structure; Oxidation-Reduction; Periodic Acid; Spectrophotometry, Ultraviolet

Ingestion of mushrooms followed after 6-12 hours by gastrointestinal symptoms and after 3-4 days by hepatic symptoms is diagnostic for the life-threatening amatoxin intoxication and should be treated as soon as possible. Four case histories are reported and recommendations for treatment are given.

Topics: Acute Kidney Injury; Adult; Amanita; Amanitins; Combined Modality Therapy; Female; Humans; Liver Function Tests; Male; Middle Aged; Mushroom Poisoning

Twenty-seven consecutive mushroom poisoning cases were followed up over a period of 14 days. Fourteen out of 27 died of liver failure. There were no deaths from renal failure. The mushrooms were identified as the amatoxin-containing Lepiota species. Therapeutic measures included nasogastric lavage, charcoal, vitamin C, vitamin B, penicillin G, corticosteroids, oral streptomycin and, in the case of a few patients, limited amounts of thioctic acid. Of the ten haemodialysed, nine died. Unfortunately charcoal haemoperfusion was not available. It appeared that therapeutic measures were ineffective and it also seemed that the amount of mushroom ingested was the determining factor for the prognosis. An important point to make is that renal failure does not occur and liver failure is always delayed (group II). For this reason all suspected cases of mushroom poisoning, regardless of absence of clinical signs and symptoms, must be hospitalised for a period of at least one week. The poisonous properties of wild mushrooms have been recognized since ancient times. However, despite awareness of their inherent dangers, serious poisoning continues to occur. Fatal intoxications can be attributed almost entirely to the amtoxin-containing species. Amanita phalloides have been blamed for over 90% of poisoning deaths in North America. There are reports of intoxications of other amatoxin-containing species in Europe, but fatalities due to Lepiota species are reported only rarely. It was previously acknowledged that the interval between ingestion of mushrooms and the onset of symptoms is longer than expected in serious poisonings.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adolescent; Adult; Amanitins; Chemical and Drug Induced Liver Injury; Child; Coma; Female; Humans; Male; Middle Aged; Mushroom Poisoning; Prognosis; Renal Dialysis; Thioctic Acid; Time Factors

The title compound 3, an amatoxin analogue containing L-alpha-aminobutyric acid instead of L-asparagine in position 1, as in natural toad stool peptides, has been synthesized. It does not inhibit the eukaryotic DNA-dependent RNA polymerase form II (or B) in concentrations up to 10(-4)M, whereas 50% inhibition is exerted in 10(-6)M solution by the corresponding Asn-analogue S-deoxo-Ile3-amaninamide 2. The striking difference seems to be due to a relatively small variation of the conformation recognized by sensitive NMR spectroscopic methods.

Topics: Amanitins; Amino Acid Sequence; Circular Dichroism; Magnetic Resonance Spectroscopy; Molecular Sequence Data; Molecular Structure; Protein Conformation; RNA Polymerase II; Temperature

The amatoxins, highly toxic components of Amanita mushrooms, strongly inhibit the DNA-dependent RNA polymerase II (or B) in eukaryotic cell nuclei. For optimal binding to the enzyme a gamma-hydroxyisoleucine side chain in the 3-position is important as in gamma-amanitin (compound 1), where the OH-group is bound in the [S]-configuration. Amanullin, a non-toxic component, having an oxygen-free isoleucine side chain no. 3, exhibits an inhibitory effect on RNA polymerase II about two orders of magnitude smaller than that of gamma-amanitin. An equal, relatively weak, inhibitory effect has previously been found with the synthetically obtained Ile3-analog 7. In the present paper the synthesis of an analog (2) bearing a gamma-hydroxyl group in the isoleucine side chain is described. The compound was found to have about the same inhibitory effect on RNA polymerase II from Drosophila embryos as amanullin and the Ile3-analog 7. Structure analysis by X-ray diffraction revealed that the hydroxyl group at the -carbon atom of side chain-3 has the [R]-configuration, the new analog thus being -deoxo[( )-hydroxy-[Ile3]-amaninamide. It follows that the [S]-configuration of this chiral center is a prerequisite to maximal toxicity. Crystallographic data demonstrating great similarity between the peptide backbones of the new analog and those of natural amatoxins are given.

Topics: Amanitins; Crystallography; Hydrogen Bonding; Molecular Structure; RNA Polymerase II; Structure-Activity Relationship

In 1986-88, 46 poisonings (4 of which were fatal) caused by amatoxin mushroom were reported. The fatalities were males aged between 7 and 65 years. They all had gastrointestinal symptoms and three of the patients died of acute hepatic insufficiency. The fourth died as a result of an intestinal perforation. The clinical and pathological alterations are compared with the findings of other authors.

Topics: Adolescent; Adult; Aged; Amanitins; Child; Humans; Male; Mushroom Poisoning; Spain

A monoclonal antibody, with high affinity against the mushroom toxin alpha-amanitin, was prepared. Administration of the Fab fragment of the monoclonal antibody to mice caused a 50-fold increase in alpha-amanitin toxicity. Electron micrographs showed normal appearance of hepatocytes but typical, amanitin-induced lesions in cells of the proximal convoluted tubules of the kidney. The pronounced nephrotoxicity is mainly explained by glomerular filtration and tubular reabsorption of the Fab-amatoxin complex and, to a lesser extent, of the immunoglobulin-amatoxin complex, which is still c. Twice as toxic as free alpha-amanitin. To our knowledge this is the first reported case where immunoglobulins or their fragments enhance rather than decrease the activity of a toxin. Accordingly, immunotherapy of Amanita mushroom poisoning in humans does not appear promising.

Topics: Amanitins; Animals; Antibodies, Monoclonal; Female; Immunoglobulin Fab Fragments; Kidney Tubules; Mice; Mice, Inbred Strains; Microscopy, Electron; Mushroom Poisoning

A novel approach for the production of [125I]amatoxin and anti-amanitin is described. The antigen and the starting material for Bolton Hunter iodination is prepared by periodic acid oxidation of the gamma-delta-dihydroxy-isoleucine side chain of alpha-amanitin followed by reductive amination. The antigen seems to be of low apparent toxicity. Chemical, spectroscopic and immunological evidence suggests that the hapten has a secondary structure similar to alpha-amanitin, but a modified tryptophanyl ring system. The establishment of a clinically useful iodinated radioimmunoassay system is possible, because the ring system seems to be of limited immunological importance.

Topics: Amanitins; Animals; Antibodies; Radioimmunoassay; Spectrophotometry, Ultraviolet; Succinimides

Topics: Amanita; Amanitins; Humans; Mushroom Poisoning

The effect of substitution of L and D amino acids in amatoxin analogues is discussed in this paper. The structure of the analog where D-alanine substitutes for glycine in position 7 has been worked out in solution by two-dimensional NMR methods using a 500 MHz instrument. The combined use of COSY and NOESY two-dimensional spectra allows a clear assignment of the resonances. The use of the coupling constants permits the calculation of the phi angles of the backbone. The NOE effects reveal the through-space contacts between protons of different peptide units, thus determining the rigidity of the amatoxin structure. On these grounds it has been possible to elucidate the conformation of the amatoxin analogue that resembles very closely that of beta-amanitin, thus explaining the high inhibitory activity toward RNA polymerase B.

Topics: Amanitins; Amino Acids; Magnetic Resonance Spectroscopy; Models, Molecular

Rabbits were immunized with fetuin-beta-amanitin. They produced amatoxin-specific immunoglobulins of various classes, predominantly IgG. The crude IgG fraction was isolated by gel filtration on Sephacryl S-300. By polyethylene glycol precipitation in the presence of tritiated amatoxin, as well as by a solid phase radioimmunoassay technique, the portion of amatoxin-specific antibodies in the IgG fraction was determined to be 5-13%. The affinity of the amatoxin-specific IgG for a tritiated amatoxin derivative was measured by equilibrium dialysis and calculated according to the method of Scatchard. The dissociation constant was 2.6 X 10(-9) M. The amatoxin-specific immunoglobulins were extracted by their affinity to alpha-amanitin-Sepharose 4B and eluted with excess alpha-amanitin. The complex was isolated in 95% yield with a ratio immunoglobulin:toxin of c. 2:1. Alternatively, the uncomplexed IgG could be eluted from the alpha-amanitin-Sepharose 4B with 5 M guanidine hydrochloride; this treatment, however, decreased the binding capacity for amatoxin by 30% (ratio 1.4:1). The purified amatoxin-specific IgG was assayed for its therapeutic efficiency in mice poisoned with alpha-amanitin, but was unable to neutralize the toxic effects of the mushroom toxin. On the contrary, equimolar amounts of the amatoxin-specific immunoglobulins enhanced the toxicity of alpha-amanitin in the mouse by a factor 2.

Topics: Amanitins; Animals; Antibodies; Chromatography, Affinity; Dialysis; Guanidine; Guanidines; Immunoglobulin G; Kinetics; Mice; Polyethylene Glycols; Rabbits; Radioimmunoassay; Spectrophotometry, Ultraviolet

In this new radioimmunoassay system for determination of amatoxins in urine and plasma, a novel chemical approach is used for antigen and 125I-tracer production, based on a detoxified alpha-amanitin derivative (aldoamanitin). Total assay time, including data processing, is less than 100 min. The lowest detectable concentration is 1 microgram/L for urine, 0.1 microgram/L for plasma. In the clinically significant range, within-run CVs are less than 8%. This new 125I-based assay is a significant improvement over existing 3H technology in terms of speed, precision, and freedom from interference.

Topics: Amanitins; Chemical Phenomena; Chemistry; Humans; Radioimmunoassay; Structure-Activity Relationship

Hepatic transport studies with amatoxins, toxic bicyclic octapeptides from poisonous mushrooms of the genus Amanita were performed, using [(6'-O,1'-N-di[3H]methyl)trp4]-alpha-amanitin and [(6'-O,1'-N-di-methyl)trp4]-[4-[3H]desmethyl)hyi3]-gamma-ama nitin. Uptake into hepatocytes from rat liver was inhibited by taurocholate and antamanide. Photoaffinity labeling studies with isolated hepatocytes and basolateral plasma membranes, using the sodium salt of (7,7-azo-3 alpha, 12 alpha-dihydroxy-5 beta-[3 beta-3H]cholan-24-oyl)-2- aminoethanesulfonic acid demonstrated that the presence of alpha-amanitin decreased the labeling of the two sinusoidal bile salt-binding membrane polypeptides with the apparent molecular weights of 54,000 and 48,000. In basolateral plasma membrane vesicles amanitin uptake was temperature-dependent and could be stimulated 1.5 to 2-fold by an out to in Na+ gradient as compared to a K+ gradient or sucrose and 2 to 2.5-fold as compared to amanitin equilibration (overshoot). Kinetic studies proved saturability of amanitin uptake in the presence and absence of a Na+ gradient. Membrane transport could be inhibited by taurocholate, antamanide, phalloidin, prednisolone, and silybin, but not by penicillin G or thioctic acid. Hepatic uptake of amatoxins is mediated by the sinusoidal bile salt-transport systems which are also involved in the uptake of antamanide and phalloidin. This supports the concept of a multispecificity of hepatic transport systems for a wide variety of amphipathic molecules.

Topics: Affinity Labels; Amanitins; Animals; Biological Transport, Active; Cell Membrane; Liver; Liver Neoplasms, Experimental; Male; Osmolar Concentration; Photochemistry; Rats; Rats, Inbred Strains

Radioactivities were measured in serum, urine, and bile of dogs at different times after intravenous injection of 14C-methyl-gamma-amanitin (14C-A) and 3H-O-methyl-dehydroxymethyl-alpha-amanitin (3H-A). For either substance, the relation between the specific plasma activity C and the time t could be best described with the function C = C1 X e- lambda 1 X t + C2 X e- lambda 2 X t. Therefore the linear open two-compartment system was selected as an adequate toxicokinetic model. Most important, the distribution volumes (in the steady state) were in the range of the extracellular space, and the total body clearances were in the range of the dog creatinine clearance. In accordance with former findings for 3H-A, 14C-A was not bound to plasma proteins. More than 80% of 14C-A was eliminated in the urine; less than 10% was found in the bile. From these data, two suggestions may be derived for the therapy of Amanita intoxication in man. First, detection in the urine of amatoxins 2 or 3 days after mushroom ingestion points to an ongoing amatoxin absorption or reabsorption from the intestine, and should lead to therapy with adsorbents and, in the absence of diarrhea, with laxatives. Second, hemoperfusion will remove significant amounts of amatoxins during the time of ongoing absorption or reabsorption and a few hours thereafter.

Topics: Amanitins; Animals; Bile; Binding Sites; Dialysis; Dogs; Fetal Blood; Half-Life; Hemoperfusion; Kinetics; Serum Albumin, Bovine

By investigations on domestic and laboratory animals great progress was achieved in the clarification of the mechanism of action and of the metabolism of mycotoxins. The aflatoxins which inhibit the synthesis of RNA in the liver and which lead to a liver necrosis are all over the world of great importance as harmful substances for man and animals. The chronic intake in subtoxic doses has a tumour-evoking effect. The amatoxins of the green Amanita phalloides are also hepatotoxic: they inhibit the synthesis of messenger RNA in the hepatocytes. The ergot alcaloids unite with alpha-adrenoceptors and evoke an inhibition of the effectiveness of beta-adrenoreceptors: they effect a vasoconstriction, a contraction of the uterus and an inhibition of the prolactin secretion. The ochratoxins have an influence on the transcription and have a nephrotoxic and teratogenic effect. The trichothezene toxins inhibit the protein synthesis and already in a low dose decrease the formation of coagulation factors and of immunoglobulins. The zearalenon has an estrogen effect: it is bound to estrogen receptors and influences the transcription in the cell nuclei. Patulin has a tumour-evoking effect. Nutrients infested by fungi are to be excluded from the consumption by man.

Topics: Aflatoxins; Amanitins; Animals; Blood Proteins; Dose-Response Relationship, Drug; Ergot Alkaloids; Humans; Kidney; Liver; Metabolic Clearance Rate; Mycotoxins; Ochratoxins; Patulin; Receptors, Adrenergic, beta; Receptors, Estrogen; Trichothecenes; Zearalenone

Four incidents of mushroom poisoning, representing four of the seven established groups of toxic mushrooms, are presented. These case reports illustrate the range of gastrointestinal and neurological symptoms caused by mushroom poisoning and reflect a nationwide increase in reports of serious poisonings in recent years. Severity of poisonings often parallels the time span between consumption and onset of symptoms, with serious poisonings having longer incubation periods. New therapies for amatoxin poisoning may reduce mortality caused by these poisonings.

Topics: Acute Kidney Injury; Adult; Aged; Amanitins; Child; Diarrhea; Female; Hallucinations; Humans; Male; Middle Aged; Mushroom Poisoning; New York; Respiratory Distress Syndrome; Thioctic Acid; Time Factors

Topics: Amanitins; Humans; Mushroom Poisoning; Pyridoxine

Topics: Adult; Amanita; Amanitins; Female; Humans; Infant, Newborn; Maternal-Fetal Exchange; Mushroom Poisoning; Pregnancy; Pregnancy Complications

Topics: Aflatoxins; Amanitins; Chemical and Drug Induced Liver Injury; Humans; Hydrocarbons, Halogenated; Liver Cirrhosis, Alcoholic; Liver Diseases; Liver Diseases, Alcoholic; Porphyrins

A fetuin derivative of alpha-amanitin was prepared and used as an antigen in rabbits. The antigen was superior to previous bovine serum albumin derivatives of beta-amanitin by its lower toxicity and high immunogenicity. On the other hand, the antibodies raised with the alpha-amanitin derivative did not show full crossreactivity with the other amatoxins, as did the immunoglobulins induced by protein derivatives of beta-amanitin. The procedure for activating nylon surfaces and coupling proteins onto them was improved with respect to surface charge and homogeneity. A partially purified IgG-fraction derived from the sera of rabbits immunized against amatoxins was covalently attached to the activated nylon surfaces. The covalently coupled immunoglobulins were complexed with a tritiated amatoxin. Then small pieces of the nylon sheet were punched out and incubated with the amatoxin solution to be analyzed. This procedure represents a method for dosing, in one step and without pipetting, the immunoglobulins and the labelled hapten. Determination of amatoxin concentrations was achieved by counting the radioactivity in the incubation fluid. The limit of detection was about 3 ng of amatoxins per ml. The radioimmunoassay was used to measure amatoxin concentrations in serum, urine, duodenal fluid, and gastric juice of patients with Amanita poisoning. Since such assays can be performed in 2-3 hr, the results can be used to determine the therapeutic protocol. The assay was likewise used to determine the concentration of amatoxins in mushroom tissue. For Amanita phalloides, for example, we found that the amatoxin concentration (mg/g dry weight) is 4.5 times higher in the gills than in the bulb.

Topics: Agaricales; Amanita; Amanitins; Animals; Antigens; Cross Reactions; Immunization; Immunoglobulins; Nylons; Rabbits; Radioimmunoassay

The preparation and characterization of a protein from wheat germ showing strong affinity to amatoxins (ABP) but differing from RNA polymerase B (II) is described. The purification, traced by [3H]amatoxin (O-methyldehydroxymethyl-alpha-amanitin), comprises 4 chromatographic steps, on Biogel A 1.5, DEAE-Sephadex, phosphocellulose, and again Biogel A 1.5. The protein exhibited in dodecyl sulfate polyacrylamide gel electrophoresis one single band with a molecular mass of 29,000 Da. Its isoelectric point is 4.9. The dissociation constant of the complex ABP-[3H]amatoxin is KD20 = 5 X 10(-7)M as determined by equilibrium dialysis against alpha-amanitin. By cyanogen bromide the protein is split into two fragments with molecular masses of 22,000 and 7,000 Da, respectively whose amino acid analyses, on summation, give the amino acid composition of ABP.

Topics: Amanitins; Amino Acids; Carrier Proteins; Molecular Weight; Plant Proteins; Plants; Triticum

Topics: Amanitins; Antidotes; Cyclopropanes; Gastric Lavage; Glutamine; Humans; Monomethylhydrazine; Muscarine; Muscimol; Mushroom Poisoning; Psilocybin

The inhibition constants (Ki) of DNA-dependent RNA polymerase B (or II) from calf thymus were measured for eight synthetically obtained bicyclic amanitin-like thioethers, two R-sulfoxides, and two S-sulfoxides. These Ki values were compared with those of alpha-amanitin, its 6'-O-methylether Ia (an R-sulfoxide), the S-sulfoxide, the sulfone, the S-deoxo derivative (Id) of Ia, and several previously described amatoxins. The necessity of a beta-methyl side chain in position 3 and a hydroxy group in proline-2 was confirmed. Additionally, the presence of an isoleucine side chain in position 6 and the absence of a side chain in position 5 was recognized as important for binding to the enzyme. In the three sulfoxide samples examined, the R-diastereomer was found to be a stronger inhibitor than the S-form. The contribution of every structural element to biological activity has been discussed.

Topics: Amanitins; Animals; Cattle; DNA-Directed RNA Polymerases; Mice; RNA Polymerase II; Structure-Activity Relationship; Thymus Gland

Topics: Amanita; Amanitins; Humans; Mushroom Poisoning

Two proteins with high affinity for amatoxins have been characterized in calf thymus nuclei, the RNA-polymerase II (or B) and a 100 K protein of unknown function. Most of the toxic effects of amatoxins are based on the inhibited synthesis of mRNA. The 100 K protein may be involved in functions of cytokinesis as suggested by experiments with PtK1 cells and a fluorescent labelled amatoxin. The molecular toxicity of phallotoxins can be understood in terms of their affinity for actin. By interaction with rabbit muscle actin the concentration of actin monomers is decreased. In hepatocytes, the phallotoxins change the structure of the microfilamentous web.

Topics: Actins; Amanitins; Animals; Cell Membrane; Chemical and Drug Induced Liver Injury; Liver; Mycotoxins; Protein Binding; Proteins; Rats; RNA Polymerase II; Subcellular Fractions

Silybin dihemisuccinate, in a concentration of 0.4 mg/ml, almost completely inhibited the uptake of an amatoxin by the perfused rat liver. Similary, silybin should also interrupt absorption of toxins due to enterohepatic circulation, e.g. in dog and man. This effect may become important in the therapy of human Amanita poisoning.

Topics: Amanitins; Animals; Depression, Chemical; Flavonoids; In Vitro Techniques; Liver; Male; Rats; Silymarin; Time Factors