Immunochemical monitoring of psilocybin and psilocin to identify hallucinogenic mushrooms.

Development of rapid and reliable immunochemical methods for monitoring psilocybin (4-phosphoryloxy-N,N-dimethyltryptamine; Pyb) and psilocin (dephosphorylated metabolite; Psi), the psychoactive compounds contained within hallucinogenic mushrooms (magic mushrooms), is desirable in order to identify these mushrooms and regulate their illicit use. Because no antibody was publicly available for this purpose, we generated two independent monoclonal antibodies (mAbs) against Pyb or Psi, and then developed enzyme-linked immunosorbent assays (ELISAs) by using them. To generate the specific antibodies, novel immunogenic conjugates were prepared by linking Pyb or Psi molecules to carrier proteins by modifying their 2-(N,N-dimethylamino)ethyl side chains. Spleen cells from mice immunized with these conjugates were fused with P3/NS1/1-Ag4-1 myeloma cells, and hybridoma clones secreting anti-Pyb and anti-Psi mAbs were established. These mAbs were characterized for their biochemical features and then applied to competitive ELISAs, which used microplates coated with Pyb or Psi linked with albumin. These ELISAs enabled the determination of Pyb or Psi with measurable ranges of ca. 0.20-20 or 0.040-2.0 µg/assay (limit of detection was 0.14 or 0.029 µg/assay), respectively. The related tryptamines were satisfactorily discriminated as exemplified by the cross-reactivity of the ELISA to determine Pyb (or Psi) with Psi (or Pyb) that were found to be 2.8 % (or <0.5 %), respectively. The Pyb and Psi contents in a dried powder of the hallucinogenic mushroom, Psilocybe cubensis, were determined to be 0.39 and 0.32 (w/w)%, respectively. The ELISAs developed using the current mAbs are promising tools for identifying illegal hallucinogenic mushrooms.

Hallucinogenic mushroom misuse reported to Texas poison centers.

BACKGROUND: Hallucinogenic mushrooms are fungi that contain the substance psilocybin. When ingested, psilocybin can induce hallucinations, euphoria, and alterations in the perception of space and time. Ingestion of hallucinogenic mushrooms may result in adverse effects. OBJECTIVE: The objective of this study was to describe hallucinogenic mushroom misuse reported to poison centers. METHODS: Cases were exposures to hallucinogenic mushrooms reported to Texas poison centers during 2000-2018 where the exposure reason was Intentional - Misuse or Intentional - Abuse (combined into the term "misuse" in the study). The distribution of the cases was determined for various factors related to patient demographics, exposure circumstances, management, and outcome. RESULTS: Texas poison centers managed 486 hallucinogenic mushroom misuse exposures. Eighty-five percent of the patients were male; the mean age was 20.9 years (range 12-57 years). The exposure route was ingestion alone in 91% of the cases. Seventy-two percent of the exposures occurred at the patient's own residence. Seventy-one percent of the patients were already at or en route to a healthcare facility when the poison center was contacted, 22% were referred to a healthcare facility by the poison center, and 7% of the patients were managed on-site. The most frequent clinical effects were neurological (72%), cardiovascular (25%), and gastrointestinal (23%). CONCLUSIONS: The majority of hallucinogenic mushroom misuse exposures involved adolescents or young adults and were male. Most of the exposures tended to be managed at a healthcare facility. The most common clinical effects tended to be neurological, cardiovascular, and gastrointestinal in nature.

Trichothecene and tremulane sesquiterpenes from a hallucinogenic mushroom Gymnopilus junonius and their cytotoxicity.

Gymnopilus junonius (Fr.) P. D. Orton (Cortinariaceae) is a hallucinogenic mushroom, a well-known poisonous mushroom that is widely known as "big laughter mushroom" because it causes excessive laughter in those who consume it. Chemical investigation of G. junonius fruiting bodies was performed, resulting in the isolation and structural identification of three sesquiterpenes (1-3), including a new trichothecene sesquiterpene (2) and a new tremulane sesquiterpene (3). Compound 1 was identified from G. junonius for the first time. The chemical structures of the new compounds were established by detailed analysis of 1D and 2D (1H-1H correlated spectroscopy [COSY], heteronuclear single quantum coherence [HSQC], and heteronuclear multiple-bond coherence [HMBC]) nuclear magnetic resonance (NMR) spectra, and high-resolution mass spectrometry (HRMS). In particular, the absolute configurations of compounds 2 and 3 were unambiguously determined by quantum chemical electronic circular dichroism (ECD) calculations. The isolated compounds (1-3) were evaluated for their cytotoxic effects on human lung and prostate cancer cell lines where trichothecene sesquiterpenes (1 and 2) showed remarkable cytotoxicity similar to that of the control drug, i.e., doxorubicin. Our findings provide experimental evidence suggesting the potential anti-cancer effects of trichothecene sesquiterpenes from a poisonous mushroom.

Polyisoprenepolyols from Fruiting Bodies of Gymnopilus spectabilis / 中国实验方剂学杂志

Objective:To study the chemical constituents, especially hallucinogenic substances of fruiting bodies of Gymnopilus spectabilis from France. Method:The fruiting bodies of G. spectabilis were extracted with chloroform-acetone-methanol (1:1:1),then the compounds were isolated and purified by silica gel column chromatography,gel chromatography,MPLC and preparative HPLC. Their structures were identified by spectroscopic analysis methods, such as 1D-NMR,2D-NMR and MS. Result:Six polyisoprenepolyols compounds were isolated and identified from fruiting bodies of G. spectabilis,namely gymnopilene A (1),hypsiziprenol B10,A10,A11(2-4),gymnopilin A10,3a(5-6). Conclusion:Compound 1 is a new compound that is named gymnopilene A,and compounds 1-4 are isolated from G. spectabilis for the first time.