Simultaneous Effects of Light Intensity and Phosphorus Supply on the Sterol Content of Phytoplankton

Sterol profiles of microalgae and their change with environmental conditions are of great interest in ecological food web research and taxonomic studies alike. Here, we investigated effects of light intensity and phosphorus supply on the sterol content of phytoplankton and assessed potential interactive effects of these important environmental factors on the sterol composition of algae. We identified sterol contents of four common phytoplankton genera, Scenedesmus, Chlamydomonas, Cryptomonas and Cyclotella, and analysed the change in sterol content with varying light intensities in both a high-phosphorus and a low-phosphorus approach. Sterol contents increased significantly with increasing light in three out of four species. Phosphorus-limitation reversed the change of sterol content with light intensity, i.e., sterol content decreased with increasing light at low phosphorus supply. Generally sterol contents were lower in low-phosphorus cultures. In conclusion, both light and phosphorus conditions strongly affect the sterol composition of algae and hence should be considered in ecological and taxonomic studies investigating the biochemical composition of algae. Data suggest a possible sterol limitation of growth and reproduction of herbivorous crustacean zooplankton during summer when high light intensities and low phosphorus supply decrease sterol contents of algae

Characterisation of a Desmosterol Reductase Involved in Phytosterol Dealkylation in the Silkworm, Bombyx mori

Most species of invertebrate animals cannot synthesise sterols de novo and many that feed on plants dealkylate phytosterols (mostly C29 and C28) yielding cholesterol (C27). The final step of this dealkylation pathway involves desmosterol reductase (DHCR24)-catalysed reduction of desmosterol to cholesterol. We now report the molecular characterisation in the silkworm, Bombyx mori, of such a desmosterol reductase involved in production of cholesterol from phytosterol, rather than in de novo synthesis of cholesterol. Phylogenomic analysis of putative desmosterol reductases revealed the occurrence of various clades that allowed for the identification of a strong reductase candidate gene in Bombyx mori (BGIBMGA 005735). Following PCR-based cloning of the cDNA (1.6 kb) and its heterologous expression in Saccharomyces cerevisae, the recombinant protein catalysed reduction of desmosterol to cholesterol in an NADH- and FAD- dependent reaction

Tolerance to drug-induced cell death favours the acquisition of multidrug resistance in Leishmania

The control of the protozoan parasite Leishmania relies on few drugs with unknown cellular targets and unclear mode of action. Several antileishmanials, however, were shown to induce apoptosis in Leishmania and this death mechanism was further studied in drug-sensitive and drug-resistant Leishmania infantum. In sensitive parasites, antimonials (SbIII), miltefosine (MF) and amphotericin B (AMB), but not paromomycin (PARO), triggered apoptotic cell death associated with reactive oxygen species (ROS). In contrast, Leishmania mutants resistant to SbIII, MF or AMB not only failed to undergo apoptosis following exposure to their respective drugs, but also were more tolerant towards apoptosis induced by other antileishmanials, provided that these killed Leishmania via ROS production. Such tolerance favored the rapid acquisition of multidrug resistance. PARO killed Leishmania in a non-apoptotic manner and failed to produce ROS. PARO resistance neither protected against drug-induced apoptosis nor provided an increased rate of acquisition of resistance to other antileishmanials. However, the PARO-resistant mutant, but not SbIII-, MF- or AMB-resistant mutants, became rapidly cross-resistant to methotrexate, a model drug also not producing ROS. Our results therefore link the mode of killing of drugs to tolerance to cell death and to a facilitated emergence of multidrug resistance. These findings may have fundamental implications in the field of chemotherapeutic interventions

Sterol composition of shellfish species commonly consumed in the United States

BACKGROUND: Shellfish can be a component of a healthy diet due to a low fat and high protein content, but the cholesterol content of some species is often cited as a reason to limit their consumption. Data on levels of non-cholesterol sterols in commonly consumed species are lacking. OBJECTIVE: Shellfish were sampled and analyzed to update sterol data in the United States Department of Agriculture (USDA) National Nutrient Database for Standard Reference. DESIGN: Using a nationwide sampling plan, raw shrimp and sea scallops, canned clams, and steamed oysters, blue crab, and lobster were sampled from 12 statistically selected supermarkets across the United States in 2007–08. For each species, four composites were analyzed, each comprised of samples from three locations; shrimp and scallops from six single locations were also analyzed separately. Using validated analytical methodology, 14 sterols were determined in total lipid extracts after saponification and derivatization to trimethylsilyethers, using gas chromatography for quantitation and mass spectrometry for confirmation of components. RESULTS: Crab, lobster, and shrimp contained significant cholesterol (96.2–27 mg/100 g); scallops and clams had the lowest concentrations (23.4–30.1 mg/100 g). Variability in cholesterol among single-location samples of shrimp was low. The major sterols in the mollusks were brassicasterol (12.6–45.6 mg/100 g) and 24-methylenecholesterol (16.7–41.9 mg/100 g), with the highest concentrations in oysters. Total non-cholesterol sterols were 46.5–75.6 mg/100 g in five single-location scallops samples, but 107 mg/100 g in the sixth, with cholesterol also higher in that sample. Other prominent non-cholesterol sterols in mollusks were 22-dehydrocholesterol, isofucosterol, clionasterol, campesterol, and 24-norcholesta-5,22-diene-3β-ol (4–21 mg/100 g). CONCLUSIONS: The presence of a wide range of sterols, including isomeric forms, in shellfish makes the analysis and quantitation of sterols in marine species more complex than in animal and plant tissues. The detailed sterol composition reported herein provides data that may be useful in research on the impact of shellfish consumption on dietary risk factors