Gastroprotective action of the ethanol extract of Leonurus sibiricus L. (Lamiaceae) in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Leonurus sibiricus L. (Lamiaceae) is a medicinal plant known in Brazil as "rubim" or "erva de macaé". It is used for various purposes, including stomach disorders. AIM OF THE STUDY: To evaluate the effect of the ethanol extract of the aerial parts of L. sibiricus (EELs) in models of gastric damage in mice. MATERIAL AND METHODS: The effect of EELs (50, 100 and 300 mg/kg, p.o., 1 h before induction) was tested on acidified ethanol (ACEt)-induced gastric ulcers. Additionally, we tested the effect of EELs (by intraduodenal administration) in the pylorus ligation (PL) model. RESULTS: Pretreatment with EELs, at 300 mg/kg, but not 50 and 100 mg/kg, reduced the relative area of gastric ulcers induced by ACEt (p < 0.01) and lipoperoxidation (p < 0.001), and increased the sulfhydryl content (p < 0.01) in the stomach in comparison with the vehicle group. Pretreatment with N-ethylmaleimide (a blocker of non-protein sulfhydryl groups, 10 mg/kg, i.p.) or glibenclamide (a KATP channel blocker, 10 mg/kg, i.p.) inhibited the gastroprotective response caused by EELs (300 mg/kg; p < 0.001), but there were no alterations due to pretreatments with inhibitors of the synthesis of prostaglandins (indomethacin, 10 mg/kg), nitric oxide (L-NAME, 70 mg/kg) or hydrogen sulfide (DL-propargylglycine, 10 mg/kg). Treatment with EELs (300 mg/kg) reduced mucus production (p < 0.001) and the volume of gastric secretion (p < 0.001) after PL without affecting gastric acidity or pH. CONCLUSIONS: These results provide evidence that EELs exerts gastroprotective action in mice, with the participation of oxidative stress and mediation of NP-SH, KATP channels and mucus production.

Stem cell-derived polarized hepatocytes.

Human stem cell-derived hepatocyte-like cells (HLCs) offer an attractive platform to study liver biology. Despite their numerous advantages, HLCs lack critical in vivo characteristics, including cell polarity. Here, we report a stem cell differentiation protocol that uses transwell filters to generate columnar polarized HLCs with clearly defined basolateral and apical membranes separated by tight junctions. We show that polarized HLCs secrete cargo directionally: Albumin, urea, and lipoproteins are secreted basolaterally, whereas bile acids are secreted apically. Further, we show that enterically transmitted hepatitis E virus (HEV) progeny particles are secreted basolaterally as quasi-enveloped particles and apically as naked virions, recapitulating essential steps of the natural infectious cycle in vivo. We also provide proof-of-concept that polarized HLCs can be used for pharmacokinetic and drug-drug interaction studies. This novel system provides a powerful tool to study hepatocyte biology, disease mechanisms, genetic variation, and drug metabolism in a more physiologically relevant setting.

Antifungal activity of the essential oil of Thymus x viciosoi against Candida, Cryptococcus, Aspergillus and dermatophyte species.

The essential oil (EO) of Thymus x viciosoi (Pau) R. Morales was isolated and analysed by GC and GC-MS. The antifungal activity of the EO and its major components against clinically relevant yeasts and molds was then measured. Their influence on the germ tube formation in Candida albicans and the influence of the EO on the metabolic function and cytoplasmic membrane integrity in the same yeast, analyzed by flow cytometry, were also studied. The EO showed high contents of carvacrol, thymol, and P-cymene. The total EO, as well as its components carvacrol and thymol, displayed very low minimum inhibitory concentrations and minimum fungicidal concentrations against all tested organisms (0.04 to 0.64 microL mL(-1)), while P-cymene showed weaker activity (2.5 to > 20.0 microL mL(-1)). They also inhibited filamentation at sub-inhibitory concentrations in C. albicans, particularly P-cymene, and the EO led to rapid metabolic arrest, disruption of the plasma membrane and consequently cell death. The EO and its main components were found to display a broad fungicidal activity through the disruption of cytoplasmic membrane integrity leading to leakage of vital intracellular compounds. In conclusion, the phenolic oil of T. x viciosoi may have potential for use in the development of clinically useful antifungal preparations.

Antifungal susceptibility testing by flow cytometry: is it the future?

The current increase in the number and significance of fungal infections, the expanding armamentarium of antifungal agents, and the emergence of the problem of antifungal drug resistance have been intensifying the importance of antifungal susceptibility testing (AST). The Clinical and Laboratory Standards Institute (CLSI, formerly NCCLS) in the United States and the Antifungal Susceptibility Testing Subcommittee of the European Committee on Antimicrobial Susceptibility Testing (AFST-EUCAST) published standard methodologies in order to achieve higher reproducibility and allow direct inter-laboratory comparison of the susceptibility results. Nevertheless, several problems remain unresolved and the methods depend on long incubation periods of a minimum of 24 h (EUCAST) or even 48 h (CLSI). Over the last 15 years, successful applications of flow cytometric techniques to AST of both yeast and moulds have been reported. These techniques are based on the analysis of a great number of fungal cells individually and frequently rely on short incubation times of no more than a few hours. Considering these attributes, flow cytometry (FC) seems to have the potential to achieve clinical usefulness in the near future. The collection of data on the reproducibility of the results and on the correlation with clinical outcomes has barely started, however. Practical validation of the experimental methodologies is not granted before a significant amount of data addressing those questions is available.