Recent studies indicated that ergosterol (Erg) helps form strongly ordered lipid domains in membranes that depend on their chemical characters. However, direct evidence of concentration-dependent interaction of Erg with lipid membranes has not been reported. We studied the Erg concentration-dependent changes in the phase behaviors of membranes using cell-sized liposomes containing 1,2-Dioleoyl-sn-glycero-3-phosphocholine (DOPC)/1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC). We observed the concentration range of phase separation in ternary membranes was significantly wider when Erg rather than cholesterol (Chol) was used as the sterol component. We used machine learning for the first time to analyze microscopic images of cell-sized liposomes and identify phase-separated structures. The automated method was successful in identifying homogeneous membranes but performance remained data-limited for the identification of phase separation domains characterized by more complex features.

The relic “the sack of Saint Francesco” has for the first time been investigated by scientific means. The sack is kept at the Franciscan Friary of Folloni near Montella in southern Italy. According to legend, the sack appeared on the doorstep of the Friary in the winter of 1224 containing bread sent from St Francesco (St Francis of Assisi), who at that time was in France. The bread was allegedly brought to the friary by an angel. We analyzed samples of the sack to obtain a radiocarbon (14C) date and to search for any remaining traces of bread. The 14C date yielded a calibrated age range of AD 1220–1295 (2σ), which places the textile in the right timeframe according to the legend. Chemical analysis by gas-chromatography with mass spectrometric detection (GC-MS) revealed the presence of ergosterol (5, 7, 22-ergostatrien-3b-ol), a known biomarker of brewing, baking, or agriculture. In this paper we have further substantiated the validity of ergosterol as a biomarker for the past presence of bread. It appears that there is a fine correspondence between the Franciscan legend and the two most decisive scientific methods relevant for analyzing the sack. Although it is not proof, our analysis shows that the sack indeed could be authentic.

Decomposition of leaf litter is an important process that releases energy and nutrients in both terrestrial and aquatic environments (Moore et al. 2004, Wallace et al. 1997); therefore, the physical, chemical and biological processes controlling leaf-litter decomposition rates can affect nutrient cycling and productivity in these systems (Cross et al. 2007, Wood et al. 2009). Several studies have shown that leaf decomposition is faster in aquatic than in terrestrial habitats due to relatively constant temperatures, continuous leaching and the physical breakdown of leaves by flowing water (Hutchens & Wallace 2002, Langhans & Tockner 2006, Langhans et al. 2008). Yet, comparatively few studies have examined these relationships in tropical systems with flooded forests. Flooding is a predominant feature of the upper Amazon Basin, but its occurrence and magnitude is complex and not strictly seasonal (Junk et al. 1989). To identify the dominant energy pathways and understand the nutrient dynamics of upper Amazon rain forests, it is imperative to investigate organic matter processing in the aquatic/terrestrial transition zones of these ecosystems.

Trypanosoma cruzi is the ethiological agent of Chagas disease. New compounds are being developed based on the biosynthesis and function of sterols, because T. cruzi has a requirement for specific endogenous sterols for growth and survival. Sterol biosynthesis inhibitors (SBIs) are drugs commonly used against fungal diseases. These drugs act by depleting essential and specific membrane components and/or inducing the accumulation of toxic intermediary or lateral products of the biosynthetic pathway. In this work we present the effects of WSP488, WSP501, and WSP561, specific inhibitors of Δ24(25)-sterol methyl transferase, on the ultrastructure of T. cruzi epimastigotes. All three drugs inhibited parasite multiplication at low concentrations, with IC50 values of 0.48, 0.44, and 0.48 μM, respectively, and induced marked morphological changes including (a) blockage of cell division; (b) swelling of the mitochondrion, with several projections and depressions; (c) swelling of the perinuclear space; (d) presence of autophagosomes and myelin-like figures; (e) enlargement of the flagellar pocket and of a cytoplasmic vacuole located in close association with the flagellar pocket; (f) detachment of the membrane of the cell body; and (g) formation of a vesicle at the surface of the parasite between the flagellar pocket and the cytostome. Our results show that these drugs are potent in vitro inhibitors of growth of T. cruzi.

Vitamin D2 from irradiated edible mushrooms might present a possible dietary source of this vitamin, subject to its bioavailability. Having previously optimized a method for the conversion of ergosterol in mushrooms to vitamin D2, this paper examines the vitamin D-enriched mushrooms (Lentinula edodes) for their bioavailability of the vitamin, using an animal model. Thirty male Wistar rats were fed for 1 week with a diet deficient in vitamin D. After this 1-week period, six rats were randomly selected and killed for analysis of initial bone mineral density, and serum level of 25-hydroxyvitamin D. A group of twelve rats of the test animals received 1 μg of vitamin D2 from irradiated mushrooms for a period of 4 weeks until being killed. The remaining twelve rats were fed un-irradiated mushrooms at the same level to act as controls. At the end of a 4-week period, the mean serum 25-hydroxyvitamin D level of the experimental group was 129·42 (sd 22·00) nmol/l whereas it was only 6·06 (sd 1·09) nmol/l in the control group. Femur bone mineral density of the experimental group of animals was significantly higher (P<0·01) than the control group. In addition, serum Ca concentrations among groups were shown to be significantly higher (P<0·01). It may be concluded from the results that vitamin D2 from UV-irradiated mushrooms is well absorbed and metabolized in this model animal system. Significant increase in femur bone mineralization (P<0·01) was shown in the presence of vitamin D2 from irradiated mushrooms compared with the controls.

The responses of the nitrophytic green algal lichen Xanthoria parietina to varying nitrogen (N) concentrations were investigated by collecting 67 X. parietina thalli from clay roof tiles from 13 sites in Portugal with different exposures to N. Concentrations of total N, chlorophyll a (a marker for the photobiont), ergosterol (a marker for the mycobiont), and thallus specific weight (TSW; thallus dry weight in relation to surface area) were quantified for each thallus to see how biont investments were related to thallus N concentrations. Thallus N ranged from 11 to 43 mg g−1 DW revealing a wider N concentration range in this lichen compared to other green algal lichen species. Both chlorophyll a and ergosterol concentrations increased with increasing thallus N, with a steeper increase of the photobiont marker. TSW was similar in all thalli without any significant effect of thallus N concentration, suggesting that thallus developmental patterns are similar in low and high thallus N concentrations. The relatively higher resource allocation to the photobiont in relation to the mycobiont with increasing thallus N concentrations is an indication of the capacity of X. parietina to meet the C demands associated with N assimilation. This result is also in agreement with the inter-specific resource allocation pattern for green algal lichens across the same N concentration range.

The aim of this investigation was to determine the NH4Cl concentration threshold, above which negative physiological effects would occur in the nitrophytic lichen Xanthoria parietina. Over a 10 month period, X. parietina thalli growing on roof tiles were exposed weekly to NH4Cl concentrations of 0·04, 0·17, 0·34 or 0·69 M. Nitrogen (N) uptake from ammonium and the concentrations of total thallus N and biont markers (chlorophyll a, ergosterol and chitin) were measured on four occasions, over the experimental period. Xanthoria parietina was able to assimilate a significant quantity of the applied ammonium. However, lichens exposed to the two higher concentrations suffered damage to both the photobiont and the mycobiont, as evidenced by reduced chlorophyll a and ergosterol concentrations, while lichens exposed to the two lower concentrations showed no significant changes in either chlorophyll a or ergosterol that could be related to the ammonium inputs. Xanthoria parietina tolerated a weekly irrigation of at least 0·17 M NH4Cl, corresponding to an N deposition of c. 1000 kg ha−1 yr−1, suggesting that this species has a very high tolerance to N pollution.

We have studied the chitin and ergosterol contents of ectomycorrhizal roots in three sets of experiments to evaluate them as indicators of fungal biomass. The first set of experiments showed that ageing had a marked effect on ergosterol concentrations. The ergosterol content of 7-month-old, brown, shrunken Pinus sylvestris L.–Paxillus involutus (Fr.) Fr. mycorrhizas was found to be only 10% of that found in white, turgid, 1- or 4-month-old specimens. This supports the hypothesis that the compound is a good indicator of living fungal biomass. Ageing had a lesser effect on chitin concentrations since the chitin levels found in 7-month-old mycorrhizas were still 60% of the levels found in 1- and 4-month-old specimens.

Consequently, the chitin[ratio ]ergosterol ratio increased from about 14 to 19 in 1- and 4-month-old mycorrhizas respectively to about 110 in 7-month-old mycorrhizas. In the second set of experiments, we found that variation in plant growth had no effect on the chitin[ratio ]ergosterol ratio in whole root systems of either Alnus incana (L.) Moench or Pinus sylvestris mycorrhizal with Paxillus involutus. In the third set of experiments, we found a constant relationship between the two marker concentrations in 10-month-old root systems of Pinus sylvestris, regardless of fungal species involved, using Paxillus involutus, Piloderma croceum Erikss. & Hjorts and Suillus variegatus (Fr.) O. Kuntze as test organisms. Taken together, the results of this study suggest that both chitin and ergosterol give reliable, but different, relative measures of fungal biomass in mycorrhizal roots. Furthermore, we demonstrate that, in combination, the two chemical markers can be used to estimate both total and living fungal biomass (derived from the chitin[ratio ]ergosterol ratio).