Skull lichens: a curious chapter in the history of phytotherapy.

Lichens growing on skulls were known in late medieval times as usnea or moss of a dead man's skull and were recommended as highly beneficial in various diseases. They were, in addition, the main ingredient of Unguentum armariun, a liniment used in a curious medical practice: the magnetic cure of wounds. We can place this chapter of the history of phytotherapy within the wider cultural context of the period, which saw the definition of nature become increasingly more fluid and open to a variety of novel interpretations.

Natural and pre-treatments induced variability in the chemical composition and morphology of lichens and mosses selected for active monitoring of airborne elements.

To enhance the reliability of the moss and lichen transplant technique for active biomonitoring of trace metals in urban environments, we evaluated the natural variability in the chemical composition of the (epilithic and epiphytic) moss Hypnum cupressiforme and the epiphytic lichen Pseudevernia furfuracea from two reference areas in NE Italy. Green shoots of epilithic mosses and lobes of epiphytic lichens from larch branches showed rather homogenous composition and were selected for the exposure in nylon bags. As different physico-chemical pre-treatments are usually applied to selected cryptogamic material before its exposure, we also evaluated the effects of oven-drying at 120 degrees C for 24h, washing in 1N HNO3 solution, and in 0.5% NH4 oxalate solution at 85 degrees C for 15 h on the chemical composition and morphology of water-washed moss shoots and lichen lobes. Pre-treatments remarkably changed the chemical composition of selected materials but not their surface morphology.

Lichen and moss bags as monitoring devices in urban areas. Part II: trace element content in living and dead biomonitors and comparison with synthetic materials.

Lobes of the lichen Pseudevernia furfuracea (L.) Zopf and shoots of the moss Hypnum cupressiforme Hedw. were subjected to different treatments (water washing, oven drying, HNO3 washing, NH4-oxalate extraction) to assess the influence of vitality on accumulation efficiency, during a 6-week exposure in bags in two Italian cities, Trieste and Naples. No trend emerged between treatments, in terms of accumulation ability, for major and trace elements. Only water-washed lichens showed an increased C and N content after exposure in both cities. Element concentrations generally reached higher values in mosses than in lichens, especially for Al, Fe, and Zn (both cities), and for Cu, Mg and Na (Naples). Surface development strongly influenced accumulation capacity of the biomonitors. Quartzose and cation exchange filters revealed, on a weight basis, a poor performance. In urban environments, surface interception of atmospheric particulate seems to play a major role in accumulation, irrespective of organism vitality.

Lichen and moss bags as monitoring devices in urban areas. Part I: influence of exposure on sample vitality.

Samples of the lichen Pseudevernia furfuracea (L.) Zopf and the moss Hypnum cupressiforme Hedw. were exposed for 6 weeks in nylon bags in two air pollution monitoring stations in Trieste and Naples (Italy) with different climates and pollution loads to evaluate influence of environmental conditions on sample vitality. This was assessed before and after exposure by transmission electron microscopy observations, K cellular location, and measurements of C, N, S and photosynthetic pigments content, CO2 gas exchange, and chlorophyll fluorescence. Almost all data sets indicate that exposures caused some damage to the species, considerably heavier in the moss, especially in Naples. The two cryptogams differed significantly in accumulation and retention of C, N, and S, the lichen clearly reflecting NO2 availability. The difference in vitality loss was related to the different ecophysiology of the species, because concentrations of phytotoxic pollutants were low during exposure. Critical notes on the analytical techniques are also given.

Oxidative stress and usnic acid content in Parmelia caperata and Parmelia soredians (Lichenes).

High light levels together with Paraquat treatment or exposure to pollutants (e.g. SO2) can cause oxidative stress in epiphytic lichens. In some cases, a significant increase in ascorbic acid and other antioxidant metabolites, sometimes occurring in lichens only, was observed. In this study, usnic acid was measured by HPLC in Parmelia caperata and Parmelia soredians thalli treated with Paraquat, a herbicide which transfers electrons from various transport systems to oxygen, producing O2*- superoxide radicals. In light, Paraquat acts by generating active oxygen species within the chloroplast, thus simulating the oxidative component of environmental stress. The significant increase in the usnic acid content measured in P. soredians thalli (+ 36.3%) is in line with the hypothesis that it has an antioxidant action, but it is in contrast to the limited increase measured in P. caperata thalli (+ 13.7%). These apparently contradictory results confirm those found in the literature, which attribute different functions to usnic acid depending on the lichen species considered and on whether they have other detoxifying substances in their thalli. These studies are of potential application-oriented interest in relation to research into new active principles to be used in the pharmaceutical, food and cosmetic fields and/or in environmental biomonitoring.

Bioaccumulation of vanadium in the epiphytic lichen Parmelia caperata from Liguria (north-west Italy).

The concentration of vanadium was measured in 35 samples of the epiphytic lichen Parmelia caperata collected along the coast of the Ligurian Sea (Italy), in order to detect the possible bioaccumulation of vanadium due to a huge crude oil burning occurred in that area in 1991. The cartographic elaboration of data shows a pattern of distribution of vanadium concentrations according to different degrees of deviation from background condition, showing that 8 years later memory of the accident is still detectable in foliose lichen thalli.

A soap technique for cell separation to study the seed coat of Sesbania punicea.

A technique is described for separating plant cells used for morphological studies. The plant material is placed in a concentrated solution of olive oil castile soap for 1-2 days or more. The material is then thoroughly washed and placed between two glass slides. The upper glass slide is lifted from the lower one, then gently pressed down several times. Through this procedure Malpighian cells of the seed coat of Sesbania punicea, mesophyll cells of Euphorbia peplus and of Trifolium pratense and cortical cells of the aerial roots of Monstera deliciosa have been separated. Various shapes of the Malpighian cells of the Sesbania punicea seed coat can be observed along with intermediates.

Membrane-bound Ca2+ distribution visualized by chlorotetracycline fluorescence during morphogenesis of soredia in a lichen.

In the lichen Parmelia caperata (L.) Ach. the distribution pattern of membrane-bound Ca2+ is investigated in the symbionts by chlorotetracycline (CTC)-induced fluorescence during the development of propagative structures, the soredia. The results demonstrate that Ca2+ accumulation in the alga and the fungus is associated with this morphogenetic process; particularly, polarized hyphal growth involves a tip-to-base Ca2+ gradient. CTC fluorescence distribution is coincident with that of cholinesterase (ChE) activity during morphogenesis of soredia. A comparison is suggested with 'embryonic ChE' of animal cells, where developmental events are regulated by a cholinergic mechanism that also modulates Ca2+ levels.

Preliminary evidence for a cholinergic-like system in lichen morphogenesis.

Membrane acetylcholinesterase activity is considered to be a marker for a cholinergic system. When temporarily expressed in differentiating cells other than the nervous or muscular ones, it may play a role in morphogenesis. In the lichen Parmelia caperata (L.) Ach., acetylcholinesterase is histochemically localized mainly in the cell walls and/or membranes of both symbionts just where they proliferate and form well-organized propagation structures, the soredia. The enzyme activity is first detected in a few algae undergoing aplanosporogenesis and later in medullary hyphae that reach the dividing algae by elongating perpendicularly to the thallus surface. This histochemical pattern that is associated with algal proliferation and oriented hyphal growth is characteristic of early morphogenesis of the soredia; when fully differentiated, they consist of an inner dividing alga and an outer hyphal envelope, both showing cholinesterase activity. Substrate specificity and inhibitor sensitivity of the histochemical staining indicate an acetylcholinesterase-like activity. However, extracts of the thallus areas where soredia develop give four bands of cholinesterase activity on disc electrophoresis: the two cathodal bands have the characteristics of acetylcholinesterase, the others of pseudocholinesterase. One of the latter hydrolyses propionylthiocholine very rapidly. The findings suggest that in lichen symbiosis, a cholinergic-like system participates in regulating morphogenetic processes such as cell division, oriented tip growth and alga-fungus membrane interactions. Environmental stimuli, particularly light, might trigger the development of soredia by modulating the activity of the cholinergic mechanism.

[Anatomical and cytohistological changes caused by aqueous extracts of Tamus communis L. leaves]. / Modificazioni anatomiche e cariologiche indotte da estratti acquosi di foglie di Tamus communis L.

Aqueous extracts of Tamus communis leaves induce root tip swelling of Medicago seedlings and disrupt the mitotic process of Allium root tips. The cells of the swollen roots of Medicago treated seedlings are much wider an shorter than the homologous control cells. In the treated roots of Allium prophases prevail and some cells contain vacuolated nuclei or micronuclei.

[Effect of aqueous extracts of Tamus communis L. leaves on the germination of seeds of various species]. / Azione di estratti acquosi di foglie di Tamus Communis L. su semi germinanti di specie diverse.

The different influence of germination and growth inhibitors in aqueous extracts, at the same concentration, of Tamus communis L. leaves, on germinating seeds and seedlings of nigra L., Lactuca sativa L., Medicago sativa L., has been shown. This action, very marked at the experimental concentrations, is not toxic, as the seeds and seedlings, if immersed in deionized water after treatments, recover their germination and growth. This recovery occurs however differently in the plants investigated.

[Histochemical localization of alcohol dehydrogenase during the aging of seeds]. / Localizzazione istochimica dell'alcool deidrogenasi durante l'invecchiamento del seme.

Alcohol dehydrogenase activity has been demonstrated in section of fresh and aged pea seeds, both air-dried and during germination. The activity in the various tissues has been observed using nitro-blue tetrazolium. From the comparison a general decrease appear in aged pea seeds and a tendency to a shift in the peaks of alcohol deydrogenase activity in the considered tissues (epidermis upper and lower, hypodermis, storage parenchyma and vascular system). The procambial zone forms the tracheary elements during germination, but in aged seeds this occur only rarely. The experiments here reported confirm the hypothesis that seed ageing includes changes in enzyme activity and in morphogenetic pathways.