Supercritical carbon dioxide extraction of the leaves and berries of Juniperus oxycedrus L. ssp. oxycedrus, coupled with a two-stage separation, allowed the entrapment of cuticular waxes in the first separator (90 bar, −10°C), thereby allowing pure essential oils to be collected in the second separator (15 bar, 10°C). An extraction carried out on a charge of leaves, at 90 bar and 50°C, gave a yield of 0.8% (w/w) with respect to the charged material, of an oil whose major constituents were germacrene D (15.9%), manoyl oxide (10.2%) and 1-epicubenol (5.4%). The berry oil obtained, at the same pressure and temperature as those for the leaves, gave a yield of 0.45% (w/w) and was composed chiefly of germacrene D (13.8%), α-pinene (10.5%) and β-myrcene (8.1%). At a constant temperature of 50°C, different extraction pressures (80, 90 and 100 bar) were tested in order to obtain the juniper wood essential oil. The extraction yield depended strongly on the extraction pressure, reaching a maximum of 14.7% (w/w) at 100 bar. The main constituents in the extract were δ-cadinene, calamenene, cubenol and 1-epi-cubenol. Hydrodistillation of the wood of J. oxycedrus gave a yield of 11.0% (w/w) of essential oil, with a reduced level of sesquiterpene hydrocarbons and an enhanced amount of oxygenated sesquiterpenes, with respect to the oil obtained by supercritical carbon dioxide extraction. The extracts obtained at different pressures were tested for cytotoxicity, antiviral and antimicrobial activities. The results showed that the extracts of leaves and berries obtained at 200 bar were cytotoxic against different cell lines used to support virus growth. As far as antiviral activiy is concerned, some of the extracts were active against a single-stranded RNA + virus (Poliovirus-1). When tested for antimicrobial activity, none of the samples were shown to be active.

Questions: What are the most relevant environmental variables influencing the distribution and the species richness of epiphytic lichens in heterogeneous areas of the Mediterranean region? What is the relative importance of substrate‐ vs. environmental‐related variables? How do climatic and disturbance factors interact? Location: The study was carried out in the Liguria region (northwestern Italy). This is a complex region where steep environmental gradients occur over a few kilometres of longitude and latitude. Methods: Data on lichen species frequencies, collected on the basis of stratified random sampling, were analyzed using non‐metric multidimensional scaling (NMS) and DCA. Indicator Value Analysis (IVA) analysis was used to find indicator species for the environmental predictors. Results: In the survey area, epiphytic lichen distribution depends on both substrate‐ and environment‐related factors. Climatic variables (in particular average yearly temperature and rainfall), anthropogenic pressures (particularly harvesting and atmospheric pollution), bark pH and texture were associated with the main axes of variability in the dataset. Mean annual rainfall is the best predictor for epiphytic lichen richness in the survey area. The particular effects of the Mediterranean region (e.g. the influence of forest fires) and of heterogeneous areas (the variability of diversity in relation to habitat) were pointed out. Several indicator species, closely linked to particular ecological conditions, were found for both substrate‐and environment‐related variables. Conclusions: The main variables involved seemed to be the same reported for other areas, but their relative importance and their spatial scale of action were in some cases different, probably due to the geomorphological and climatic heterogeneity of the survey area. The significant influence of climatic predictors and disturbance factors on epiphytic lichens was confirmed, suggesting that these organisms may be used successfully to describe ecological trends in natural environments, also providing practical information at ecosystem level.

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Progressive multifocal leukoencephalopathy (PML) is a demyelinating disease emerging as a complication of some novel monoclonal antibody therapies. Disease risk and severity show a wide variability, with fatality ranging from 30% to 90%, probably depending on many risk determinants as demonstrated for natalizumab-associated PML. Rituximab therapy is used to treat hematological malignancies and autoimmune disease, but little is known about PML risk stratification and management. We describe a clinical case with very early diagnosis, extensive treatment, but a fatal outcome, highlighting the need for future studies aimed to better understand PML risk.