infekce a stres v raném věku během zranitelného období vývoje cns souvisejí se zvýšeným rizikem vzniku neuropsychiatrických poruch, jako je schizofrenie. na základě teorie dvojího zásahu u schizofrenie je hypoteticky ke spuštění poruchy nutný ještě druhý inzult rùzné modality. neonatální aplikace lipopolysacharidu (lPs, známého také jako endotoxin) u potkanù je uznávaným modelem vedoucím k behaviorálnímu deficitu a neurokognitivním poruchám na podkladě imunitní stimulace v raném věku a používá se jako neurovývojový model psychózy. V této práci jsme na modelu potkana studovali, jak neonatální podání lPs spolu s dysbiózou vyvolanou chronickou léčbou antibiotiky jako druhým zásahem na základě teorie dvojího zásahu ovlivní metabolické dráhy pomocí necílené metabolomické analýzy. hodnotili jsme rozdíly mezi metabolomy kontrolní a testovací skupiny (vystavené lPs a/nebo léčbě antibiotiky), abychom zjistili rozdíly mezi jejich metabolitovými profily. Vzorky skupiny léčené lPs vykazovaly pozoruhodné metabolomické změny ve srovnání s kontrolními subjekty. Jako nejvíce ovlivněné divergentní dráhy byly určeny dráhy pyrimidinového metabolismu a dráhy biosyntézy pantothenátu a koenzymu a (coa); p-hodnota upravená podle gama je 0,04275 a 0,04278. Léčba ATB nevedla k žádným změnám metabolomu, a naopak maskovala účinky vyvolané aplikací lPs. S odkazem na tyto údaje jsme schopni naznačit, že neonatální výzva LPS hraje významnou roli v rozkladu pyrimidinového metabolismu a drah biosyntézy pantothenátu a coa, které by mohly být identifikovány jako metabolické dráhy zapojené do patogeneze schizofrenie. Vzhledem k pøedběžné povaze naší studie je nutné naše zjištění ověøit v budoucím výzkumu schizofrenie.
infections and stress in early life, during a vulnerable development period of the cns, are related to an increased risk of the development of neuropsychiatric disorders such as schizophrenia. Based on the dual-intervention theory of schizophrenia, a second insult modality is hypothesized to be required to trigger the disorder. neonatal lipopolysaccharide (lPs, also known as endotoxin) treatment in rats is an acknowledged model for inducing the behavioral deficits and neurocognitive impairments caused by an early-life immune challenge and is used as a neurodevelopmental model of psychosis. in the present work, we studied in the rat model how the neonatal lPs administration along with dysbiosis induced by chronic antibiotic treatment as a second hit based on the dual hit theory will affect metabolic pathways by an untargeted metabolomic analysis. We have evaluated differences between the metabolome of control and test groups (exposed to lPs and/or antibiotic treatment) to determine differences between their metabolite profiles. the lPs-treated group samples have exhibited notable metabolomic changes compared to the control subjects. the most affected divergent pathways were determined as Pyrimidine metabolism and Pantothenate and coenzyme a (coa) biosynthesis pathways (the gamma-adjusted p-values are 0.04275 and 0.04278, respectively). the atB treatment did not result in any changes in metabolome and, on the contrary, masked the effects induced by lPs treatment. referring to these data, we are able to suggest that a neonatal lPs challenge plays a significant role in the discomposing Pyrimidine metabolism and Pantothenate and coa biosynthesis pathways, which could be identified as metabolism pathways involved in schizophrenia pathogenesis. Due to the preliminary nature of our study, it is necessary to confirm our findings in future schizophrenia research
Compounds from the N-benzylphenethylamine (NBPEA) class of novel psychoactive substances are being increasingly utilized in neurobiological and clinical research, as diagnostic tools, or for recreational purposes. To understand the pharmacology, safety, or potential toxicity of these substances, elucidating their metabolic fate is therefore of the utmost interest. Several studies on NBPEA metabolism have emerged, but scarce information about substances with a tetrahydrobenzodifuran ("Fly") moiety is available. Here, we investigated the metabolism of 2-(8-bromo-2,3,6,7-tetrahydrobenzo[1,2-b:4,5-b']difuran-4-yl)-N-(2-methoxybenzyl)ethan-1-amine (2C-B-Fly-NBOMe) in three different systems: isolated human liver microsomes, Cunninghamella elegans mycelium, and in rats in vivo. Phase I and II metabolites of 2C-B-Fly-NBOMe were first detected in an untargeted screening and identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Several hypothesized metabolites were then synthesized as reference standards; knowledge of their fragmentation patterns was utilized for confirmation or tentative identification of isomers. Altogether, thirty-five phase I and nine phase II 2C-B-Fly-NBOMe metabolites were detected. Major detected metabolic pathways were mono- and poly-hydroxylation, O-demethylation, oxidative debromination, and to a lesser extent also N-demethoxybenzylation, followed by glucuronidation and/or N-acetylation. Differences were observed for the three used media. The highest number of metabolites and at highest concentration were found in human liver microsomes. In vivo metabolites detected from rat urine included two poly-hydroxylated metabolites found only in this media. Mycelium matrix contained several dehydrogenated, N-oxygenated, and dibrominated metabolites.
- Publikační typ
- časopisecké články MeSH
BACKGROUND: Tuberous sclerosis complex (TSC), a multi-system genetic disorder often associated with autism spectrum disorder (ASD), is caused by mutations of TSC1 or TSC2, which lead to constitutive overactivation of mammalian target of rapamycin (mTOR). In several Tsc1+/- and Tsc2+/- animal models, cognitive and social behavior deficits were reversed by mTOR inhibitors. However, phase II studies have not shown amelioration of ASD and cognitive deficits in individuals with TSC during mTOR inhibitor therapy. We asked here if developmental epilepsy, common in the majority of individuals with TSC but absent in most animal models, could explain the discrepancy. METHODS: At postnatal day P12, developmental status epilepticus (DSE) was induced in male Tsc2+/- (Eker) and wild-type rats, establishing four experimental groups including controls. In adult animals (n = 36), the behavior was assessed in the paradigms of social interaction test, elevated plus-maze, light-dark test, Y-maze, and novel object recognition. The testing was carried out before medication (T1), during a 2-week treatment with the mTOR inhibitor everolimus (T2) and after an 8-week washing-out (T3). Electroencephalographic (EEG) activity was recorded in a separate set of animals (n = 18). RESULTS: Both Tsc2+/- mutation and DSE caused social behavior deficits and epileptiform EEG abnormalities (T1). Everolimus led to a persistent improvement of the social deficit induced by Tsc2+/-, while deficits related to DSE did not respond to everolimus (T2, T3). CONCLUSIONS: These findings may contribute to an explanation why ASD symptoms in individuals with TSC, where comorbid early-onset epilepsy is common, were not reliably ameliorated by mTOR inhibitors in clinical studies.
- MeSH
- autistická porucha * MeSH
- haploinsuficience MeSH
- krysa rodu rattus MeSH
- status epilepticus * MeSH
- TOR serin-threoninkinasy genetika MeSH
- tuberin genetika MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
N-Benzylphenethylamines are novel psychedelic substances increasingly used for research, diagnostic, or recreational purposes. To date, only a few metabolism studies have been conducted for N-2-methoxybenzylated compounds (NBOMes). Thus, the available 2,5-dimethoxy-4-(2-((2-methoxybenzyl)amino)ethyl)benzonitrile (25CN-NBOMe) metabolism data are limited. Herein, we investigated the metabolic profile of 25CN-NBOMe in vivo in rats and in vitro in Cunninghamella elegans (C. elegans) mycelium and human liver microsomes. Phase I and phase II metabolites were first detected in an untargeted screening, followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) identification of the most abundant metabolites by comparison with in-house synthesized reference materials. The major metabolic pathways described within this study (mono- and bis-O-demethylation, hydroxylation at different positions, and combinations thereof, followed by the glucuronidation, sulfation, and/or N-acetylation of primary metabolites) generally correspond to the results of previously reported metabolism of several other NBOMes. The cyano functional group was either hydrolyzed to the respective amide or carboxylic acid or remained untouched. Differences between species should be taken into account in studies of the metabolism of novel substances.
- Publikační typ
- časopisecké články MeSH
In recent years, the use of synthetic cannabinoids (SCs) as drugs of abuse has greatly increased. SCs are associated with a risk of severe poisoning or even death. Therefore, more rapid, cost effective and reliable methods are needed, especially for the screening of drivers after traffic accidents and for detailed toxicological analysis in forensic laboratories. In this study, we developed a lateral flow immunoassay (LFIA) and an enzyme linked immunosorbent assay (ELISA) for the detection of JWH-200 in oral fluids. For this purpose a new hapten was prepared using a ten-step synthetic route. The developed immuno methods are based on antibodies obtained from rabbit immunized with synthesized hapten conjugated to carrier protein. The proposed methods are highly sensitive (LODLFIA = 0.08 ± 0.04 ng mL-1; LODELISA = 0.04 ± 0.02 ng mL-1). They were applied to the quantification of JHW-200 in spiked oral fluids. The recoveries ranged from 82 to 134% for both methods. The results correlated excellently with results obtained using UHPLC-MS/MS (R2LFIA = 0.99; R2ELISA = 0.99). Our developed methods could be an important tool for analyses of JWH-200 in human oral fluids. The one-step LFIA is particularly suitable for roadside and on-site monitoring due to the rapid qualitative results it delivers, while the ELISA is especially useful for laboratory quantitative analyses of positive samples captured by LFIA.
- Publikační typ
- časopisecké články MeSH
INTRODUCTION: The use of new psychoactive substances as drugs of abuse has dramatically increased over the last years. Hallucinogenic phenethylamines gained particular popularity as they have both stimulating and psychedelic effects. Although generally perceived as safe, these illicit drugs pose a serious health risk; they have been linked to cases of severe poisoning or even deaths. Therefore, simple, cost-effective and reliable methods are needed for rapid determination of abused hallucinogens. METHODS: For this purpose, two haptens derived from 2C-H were designed, synthesized and subsequently attached to a carrier protein. Polyclonal antibodies obtained from a rabbit immunized with one of the prepared immunogens were used for the development of two immunoassays. RESULTS: In this study, a lateral flow immunoassay (LFIA) and an enzyme linked immunosorbent assay (ELISA) for the detection of 2C-B and related hallucinogenic phenethylamines in urine were developed. The presented LFIA is primarily suitable for on-site monitoring as it is simple and can provide a visual evidence of 2C-B presence within a few minutes. Its reasonable sensitivity (LODLFIA = 15 ± 7 ng mL-1) allows detection of the drug presence in urine after acute exposure. For greater accuracy, highly sensitive ELISA (LODELISA = 6 ± 3 pg mL-1) is proposed for toxicological quantitative analyses of positive samples captured by the LFIA. DISCUSSION: The comparison of the ELISA with the well-established UHPLC-MS-MS method shows excellent agreement of results, which confirms good potential of the ELISA to be used for routine analyses of 2C-B and related hallucinogenic phenethylamines of both main sub-families.
- MeSH
- dimethoxyfenylethylamin analogy a deriváty chemie imunologie moč MeSH
- halucinogeny chemie imunologie moč MeSH
- hapteny chemie imunologie MeSH
- imunoanalýza ekonomika metody MeSH
- lidé MeSH
- odhalování abúzu drog přístrojové vybavení metody MeSH
- reprodukovatelnost výsledků MeSH
- tandemová hmotnostní spektrometrie přístrojové vybavení metody MeSH
- zakázané drogy chemie imunologie moč MeSH
- zdraví dobrovolníci pro lékařské studie MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- validační studie MeSH
