Antifungal compounds from cultures of dairy propionibacteria type strains

Antifungal compounds from cultures of five type strains of dairy propionibacteria, as well as from the cultivation medium, were studied. Cell-free supernatants and medium were fractionated by C(18) solid phase extraction. The aqueous 95% acetonitrile fractions were analyzed by GC-MS or subjected to reversed-phase HPLC, to identify, quantify or isolate antifungal substances. The resulting HPLC fractions were screened for antifungal activity against the mold Aspergillus fumigatus and the yeast Rhodotorula mucilaginosa. Active fractions were further separated by HPLC and the structures of the compounds were determined by spectroscopic and chromatographic methods. All five strains produced 3-phenyllactic acid, at concentrations ranging from 1.0 microg mL(-1) (Propionibacterium freudenreichii ssp. shermanii) to 15.1 microg mL(-1) (Propionibacterium thoenii), and at L/D -ratios ranging from 2 : 3 (Propionibacterium acidipropionici) to 9 : 1 (Propionibacterium freudenreichii). A number of active compounds found in cultures of propionibacteria were also present in noninoculated growth medium: two antifungal diketopiperazines, cyclo(L-Phe-L-Pro) and cyclo(L-Ile-L-Pro), and seven antifungal linear peptides. Three of the linear peptides corresponded to sequences found in the medium component casein, suggesting their origin from this component, whereas the diketopiperazines were suggested to be formed from medium peptides by heat treatment.     

Beta-cell PDE3B regulates Ca2+-stimulated exocytosis of insulin

cAMP signaling is important for the regulation of insulin secretion in pancreatic beta-cells. The level of intracellular cAMP is controlled through its production by adenylyl cyclases and its breakdown by cyclic nucleotide phosphodiesterases (PDEs). We have previously shown that PDE3B is involved in the regulation of nutrient-stimulated insulin secretion. Here, aiming at getting deeper functional insights, we have examined the role of PDE3B in the two phases of insulin secretion as well as its localization in the beta-cell. Depolarization-induced insulin secretion was assessed and in models where PDE3B was overexpressed [islets from transgenic RIP-PDE3B/7 mice and adenovirally (AdPDE3B) infected INS-1 (832/13) cells], the first phase of insulin secretion, occurring in response to stimulation with high K(+) for 5 min, was significantly reduced ( approximately 25% compared to controls). In contrast, in islets from PDE3B(-/-) mice the response to high K(+) was increased. Further, stimulation of isolated beta-cells from RIP-PDE3B/7 islets, using successive trains of voltage-clamped depolarizations, resulted in reduced Ca(2+)-triggered first phase exocytotic response as well as reduced granule mobilization-dependent second phase, compared to wild-type beta-cells. Using sub-cellular fractionation, confocal microscopy and transmission electron microscopy of isolated mouse islets and INS-1 (832/13) cells, we show that endogenous and overexpressed PDE3B is localized to insulin granules and plasma membrane. We conclude that PDE3B, through hydrolysis of cAMP in pools regulated by Ca(2+), plays a regulatory role in depolarization-induced insulin secretion and that the enzyme is associated with the exocytotic machinery in beta-cells.     

Effect of Mn toxicity on morphological and physiological changes in two <Emphasis Type="Italic">Populus cathayana</Emphasis> populations originating from different habitats

The cuttings of Populus cathayana were exposed to four different manganese (Mn) concentrations (0, 0.1, 0.5 and 1 mM) in a greenhouse to investigate the toxicity of Mn and the detoxifying responses of woody plants. Two contrasting populations of P. cathayana, which were from wet and dry climate regions in western China, respectively, were examined in our study. The results showed that high concentration of Mn caused significant decrease in shoot height, biomass accumulation, and leaf number and leaf areas. Injuries to the anatomical features of leaves were also found as the reduced thickness of palisade and spongy parenchyma, the decreased density in the conducting tissue and the collapse and split in the meristematic tissue in the central vein. Moreover, Mn treatments caused the accumulation of hydrogen peroxide (H₂O₂), and then resulted in oxidative stress indicated by the oxidation of proteins and DNA. Many physiological responses were employed to cope with the toxicity of Mn, including the increase in the contents of non-protein thiol (NP-SH), phytochelatins (PCs) and phenolics compounds and the stimulated activities of guaiacol peroxidase (GPX) and polyphenol oxidase (PPO) for the chelation of Mn and for the antioxidation of reactive oxygen species. The population from dry climate habitat showed a lower leaf concentration of Mn, higher contents of the chelators, and higher activities of GPX and PPO than did the wet climate population at the same Mn treatment, thereby possessing a superior Mn tolerance. In both populations, most of the Mn was accumulated in the shoot, which is favorable regarding phytoremediation.     

Genetic diversity and differential in vitro responses to Ni in <Emphasis Type="Italic">Cenococcum geophilum</Emphasis> isolates from serpentine soils in Portugal

Amplified fragment length polymorphism (AFLP) analysis was used to investigate the genetic diversity in isolates of the ectomycorrhizal fungus Cenococcum geophilum from serpentine and non-serpentine soils in Portugal. A high degree of genetic diversity was found among C. geophilum isolates; AFLP fingerprints showed that all the isolates were genetically distinct. We also assessed the in vitro Ni sensitivity in three serpentine isolates and one non-serpentine isolate. Only the non-serpentine isolate was significantly affected by the addition of Ni to the growth medium. At 30 microg g(-1) Ni, radial growth rate and biomass accumulation decreased to 73.3 and 71.6% of control, respectively, a highly significant inhibitory effect. Nickel at this concentration had no significant inhibitory effect on serpentine isolates, and so the fitness of serpentine isolates, as evaluated by radial growth rate and biomass yield, is likely unaffected by Ni in the field. In all isolates, the Ni concentration in the mycelia increased with increasing Ni concentration in the growth medium, but two profiles of Ni accumulation were identified. One serpentine isolate showed a linear trend of Ni accumulation. At the highest Ni exposure, the concentration of Ni in the mycelium of this isolate was in the hyperaccumulation range for Ni as defined for higher plants. In the remaining isolates, Ni accumulation was less pronounced and seems to approach a plateau at 30 microg g(-1) Ni. Because two profiles of Ni accumulation emerged among our Ni-insensitive serpentine isolates, this result suggests that different Ni detoxification pathways may be operating. The non-serpentine isolate whose growth was significantly affected by Ni was separated from the other isolates in the genetic analysis, suggesting a genetic basis for the Ni-sensitivity trait. This hypothesis is further supported by the fact that all isolates were maintained on medium without added Ni to avoid carry-over effects. However, because AFLP analysis failed to distinguish between serpentine and non-serpentine isolates, we cannot conclude that Ni insensitivity among our serpentine isolates is due to evolutionary adaptation. Screening a larger number of isolates, from different geographical origins and environments, should clarify the relationships between genetic diversity, morphology, and physiology in this important species.     

Visual pigments in a living fossil,the Australian lungfish <Emphasis Type="Italic">Neoceratodus forsteri</Emphasis>

Background  

One of the greatest challenges facing the early land vertebrates was the need to effectively interpret a terrestrial environment. Interpretation was based on ocular adaptations evolved for an aquatic environment millions of years earlier. The Australian lungfish Neoceratodus forsteri is thought to be the closest living relative to the first terrestrial vertebrate, and yet nothing is known about the visual pigments present in lungfish or the early tetrapods.     

Mechanistic studies of electrophilic protease inhibitors of full length hepatic C virus (HCV) NS3

The inhibition mechanism of electrophilic peptide-based protease inhibitors of full-length hepatitis C virus (HCV) NS3 has been investigated by determining the K(i)-values for a series of compounds differing in the electrophilicity and acidity of the C-terminal residue at pH-values above and below the pK(a) of the catalytic histidine (6.85) and at two different ionic strengths. Electrophilic compounds with a pentafluoroethyl ketone group showed stronger inhibition at pH 8 than pH 6, as expected for a mechanism requiring an unprotonated catalytic histidine. However, the difference was only significant at high ionic strength. In contrast, electrophilic compounds with an acidic C-terminal group or a cyclic P1 residue showed a lower inhibitory effect at pH 8 than at pH 6, inconsistent with a mechanism-based inhibition. Moreover, all electrophilic compounds had an unexpectedly strong inhibition at pH 6, when mechanism-based inhibition is unlikely. The results suggest that for some of the electrophilic compounds the reactive group may not be properly positioned in the active site and that binding of these inhibitors is a result of non-covalent interactions. The nature of these interactions is discussed.     

Occurrence of Lepidopterism caused by the moth Hylesia nigricans (Berg) (Lepidoptera: Saturniidae) in Rio Grande do Sul State, Brazil

Lepidopterism by Hylesia nigricans (Berg) moth is recorded for the first time in southern Brazil. Preventive strategies of control are proposed based on information on the biology and ecology of this moth.     

From the saltpan to the plate: An evaluation of the use of the edible halophyte Salicornia ramosissima in catering

The halophyte species Salicornia ramosissima has long been consumed as food and is currently regarded as a high-value gourmet item. However, given that these plants grow in estuarine areas, often impacted by pollution, and are prone to the accumulation of ions, the risks associated with food borne pathogens or toxic metals has to be carefully considered. The objective of this work was to gather evidence that S. ramossissima harvested locally in an estuarine system (Ria de Aveiro, Portugal) can be safely consumed as a healthy and catering-amenable food. Fresh shoots, taken as green vegetables, and dried powdered shoots (green salt) intended as seasoning, were analysed for total aerobic bacteria, total coliforms, Escherichia coli, Bacillus cereus and fungi (yeasts and moulds). The nutritional profile and the concentration of macro and micro elements, including potential toxic elements, was also established. Fresh shoots and green salt were incorporated in catering meals (soup and a meat dish) and subjected to sensory analyses. The microbiological quality of fresh shoots and green salt was compliant with the guidelines for salad greens and herbs and spices, respectively. The nutritional and elemental profile revealed that fresh shoots have a high content in fibre and NaCl. From the concentration of Na determined in green salt, a daily dose of 7 g was estimated as corresponding to the recommended daily dietary intake Na. None of the other elements was present in concentrations that could exceed the dietary intake limits. In the sensory tests, fresh shoots were well accepted as salad greens but green salt, used as seasoning, was considered as insufficient in terms of saltiness and does not completely substitute cooking salt. The results confirm the nutritional and sensory value of S. ramossisima and represent a promising prospect for the incorporation of locally harvested or crop cultivated plants in catering meals.     

The role of organic matter and microbial community controlling nitrate reduction under elevated ferrous iron concentrations in boreal lake sediments

Hydrobiologia - The nitrogen availability, that affects the greenhouse gas emission and the trophic level of lakes, is controlled mainly by microbial processes. We measured in a boreal nitrate and...