Assessment of the possible health risks associated with the consumption of botanical preparations of Mitragyna speciosa (kratom).

The current report summarises the work performed in the context of the European Food Risk Assessment Fellowship Programme (EU-FORA), which included the evaluation of health risks associated with the consumption of botanical preparations of Mitragyna speciosa (kratom). Mitragyna speciosa is a tree native to Southeast Asia, where its leaves and preparations of the leaves have been used for centuries, among others, as a stimulant or as a traditional herbal medicine. Preparations of the plant have recently gained increasing popularity in other parts of the world, and are presently also accessible via online platforms, e.g. as food supplements. Kratom has been considered a botanical of possible health concern by the FDA and EFSA, which together with its increasing popularity, makes kratom a subject of international concern. Major alkaloids of the plant, mitragynine and 7-hydroxymitragynine, are agonists of the µ-opioid human receptor and are assumed to be mainly responsible for its psychoactive effects. The aim of the present project was to conduct an assessment of potential health risks associated with oral use of kratom-based preparations. The animal and human data that were evaluated in the course of the current assessment indicate that kratom consumption has the potential to not only lead to adverse neurological effects, including addiction and withdrawal syndrome, but also to elicit distinct organ toxicity with respect to e. g. liver and kidney as target organs. Nevertheless, actual risk characterisation is impeded by considerable uncertainties. Such uncertainties, based on the variability in composition of kratom preparations, insufficient information on dose-response relationships and on limited data on long-term use effects, currently do not allow the derivation of distinct health based guidance values for kratom/kratom preparations. Further information from well-designed studies, conducted with kratom preparations that have been clearly defined with respect to their composition, would be required to enable a more refined risk assessment of this botanical.

Induction of EpRE-mediated gene expression by a series of mediterranean botanicals and their constituents.

ETHNOPHARMACOLOGICAL RELEVANCE: A variety of Mediterranean plant species, traditionally used for the prevention and treatment of several health conditions, contain ingredients with potential biological activity of which many remain unexplored. Among the beneficial health effects of bioactive phytochemicals is the activation of cellular defense mechanisms involving the activation of EpRE (electrophile responsive element) - mediated changes in gene expression. AIM OF THE STUDY: The present study aimed to identify botanicals and their active constituents able to activate the EpRE mediated gene expression within a series of Mediterranean plant species known for their hepatoprotective and/or cardioprotective properties. MATERIALS AND METHODS: Methanolic extracts of 18 botanicals were prepared and tested for their ability to induce gene expression in EpRE-LUX reporter cells. Subsequently, LC-MS (Liquid Chromatography Mass Spectrometry) analysis combined with MAGMa (MS Annotation based on in silico Generated Metabolites) software for automated compound annotation was used to facilitate tentative identification of the active constituents within two of the active extracts. Selected annotated compounds were tested in the EpRE-LUX reporter gene assay followed by definite identification of the most active ones. RESULTS: It appeared that 9 of the 18 extracts were able to activate EpRE-mediated gene expression. Many active ingredients of the methanolic extracts from Juglans regia and Rhamnus frangula were revealed. Among them, chrysophanol and aloe-emodin were confirmed to be active EpRE inducing ingredients and were definitely identified in the Rhamnus Frangula extract. CONCLUSIONS: The protective effect of half of the tested botanical varieties via the activation of EpRE-mediated gene expression was confirmed. The study also provided an example of how in vitro bioassays can be combined with LC-MS and the automated chemical annotation software MAGMa, to identify biologically active constituents in complex botanical extracts.

In vitro bioassays to evaluate beneficial and adverse health effects of botanicals: promises and pitfalls.

This review provides an update on the promises and pitfalls when using in vitro bioassays to evaluate beneficial and adverse health effects of botanicals and botanical preparations. Important issues addressed in the paper are: (i) the type of assays and biological effects available; (ii) false-positives, false-negatives and confounding factors; (iii) matrix and combination effects; (iv) extrapolation of in vitro data to the in vivo situation; (v) when (not) to use bioassays; and (vi) identification of active constituents. It is concluded that in vitro bioassays provide models to detect beneficial as well as adverse activities, but that linking these observations to individual ingredients and extrapolations to the in vivo situation is more complicated than generally anticipated.

Growth kinetic analysis of bi-recombinant poliovirus vaccine strains.

Attenuated strains of Sabin poliovirus vaccine replicate in the human gut and in rare cases may cause vaccine-associated paralytic poliomyelitis (VAPP). Mutations at specific sites of the genome and recombination between Sabin strains may result in the loss of the attenuated phenotype of OPV (Oral Poliovirus Vaccine) strains and the acquisition of traits characteristic of wild polioviruses, such as increased neurovirulence and loss of temperature sensitivity. In this study, we determined the phenotypic traits such as temperature sensitivity and growth kinetics of eight OPV isolates (six bi-recombinant and two non-recombinant). The growth phenotype of each isolate as well as of Sabin vaccine strains in Hep2 cell line at two different temperatures (37 and 40 degrees C) was evaluated using two different assays, RCT test (Reproductive Capacity at different Temperatures) and one-step growth curve analysis. Moreover, the nucleotide and amino acid positions in the genomes of the isolates that have been identified as being involved in the attenuated and thermo sensitive phenotype of Sabin vaccine strains were investigated. Mutations that result in loss of the attenuated and thermo sensitive phenotype of Sabin vaccine strains were identified in the genomes of all isolates. Both mutations and recombination events correlated well with the reverted phenotypic traits of OPV-derivatives. In the post-eradication era of wild polioviruses, the identification and the characterization (genomic and phenotypic) of vaccine-derived polioviruses become increasingly important in order to prevent cases or even outbreaks of paralytic poliomyelitis caused by neurovirulent strains.