Effect of different forms of alkali treatment on specific fermentation inhibitors and on the fermentability of lignocellulose hydrolysates for production of fuel ethanol

Treatment with alkali, particularly overliming, has been widely used as a method for the detoxification of lignocellulose hydrolysates prior to ethanolic fermentation. However, the mechanisms behind the detoxification effect and the influence of the choice of cation have not been well understood. In this study, a dilute acid hydrolysate of spruce and an inhibitor cocktail consisting of six known inhibitors were used to investigate different alkali detoxification methods. The various treatments included the addition of calcium hydroxide, sodium hydroxide, potassium hydroxide, and ammonia to pH 10.0 and subsequent adjustment of the pH to 5.5 with either sulfuric or hydrochloric acid as well as treatment with the corresponding amounts of calcium, sodium, and potassium as sulfate or chloride salts at pH 5.5. An RP-HPLC method was developed for the separation of 18 different inhibitors in the hydrolysate, including furaldehydes and phenolics. Detection and quantification were carried out by means of UV, DAD, and ESI-MS in negative mode. Treatment of the spruce hydrolysate with alkali resulted in up to approximately 40% decrease in the concentration of furaldehydes. The effects on the aromatic compounds were complex. Furthermore, SFE was performed on the precipitate formed during alkali treatment to evaluate the inhibitor content of the precipitate, and the following RP-HPLC analysis implied that potential inhibitors were removed mainly through conversion rather than through filtration of precipitate. Parallel experiments in which sulfuric acid or hydrochloric acid was used for acidification to pH 5.5 after alkali treatment indicated that the choice of anion did not affect the removal of inhibitors. Detoxification with calcium hydroxide and ammonia resulted in better fermentability using Saccharomyces cerevisiae than detoxification with sodium hydroxide. The results from the experiments with the inhibitor cocktail indicated that the positive effects of alkali treatment are difficult to explain by removal of the inhibitors only and that possible stimulatory effects on the fermenting organism warrant further attention.     

Aluminium impact on respiration of lotic mayflies at low pH

Three species of stream-living mayfly nymphs were exposed to total inorganic (labile) monomeric aluminium levels of 0, 0.5, and 2.0 mg L^?1 at pH 4.0 and 4.8 for 10 days. Oxygen consumption rate of the nymphs was measured, using a closed-cylinder technique and micro-Winkler titration. The animals showed significant increases in respiration with raised Al concentrations. The mayfly species Ephemera danica, which is restricted to the less heavily acidified region of South Sweden, was most affected by Al. The response of the two Heptagenia species to elevated Al was less pronounced. The effect of pH, however, seemed less important. Two possible reasons for the respiration increase at high Al levels are: (1) A “chemical impact route”, consisting of a compensatory mechanism due to decreased O₂ transport efficiency, because of impaired osmoregulation and ion transport; and/or (2) a “mechanical impact route” with Al hydroxide precipitation and mucus formation on the gills, causing lowered respiration efficiency. This stress of the increased respiration rate costs energy. Thus less will be available for growth and reproduction. A model of the proposed impact routes is presented.     

Contrasting effects of puerarin and daidzin on glucose homeostasis in mice

Puerarin and daidzin are the major isoflavone glucosides found in kudzu dietary supplements. In this study, we demonstrated that puerarin significantly improves glucose tolerance in C57BL/6J-ob/ob mice, an animal model of type 2 diabetes mellitus, blunting the rise in blood glucose levels after i.p. administration of glucose. In contrast, daidzin, the O-glucoside, had a significant but opposite effect, impairing glucose tolerance as compared to saline-treated controls. When they were administered i.p. with (14)C-glucose to C57BL/6J lean mice, puerarin inhibited glucose uptake into tissues and incorporation into glycogen, while daidzin stimulated glucose uptake, showing an opposite effect to puerarin. Puerarin also antagonized the stimulatory effect of decyl-beta-D-thiomaltoside, an artificial primer of glycogen synthesis, which increases (14)C-glucose uptake and incorporation into glycogen in mouse liver and heart. A liquid chromatography-tandem mass spectrometry procedure was used to investigate the metabolism and bioavailability of puerarin and daidzin. The blood puerarin concentration-time curve by i.p. and oral administration indicated that puerarin was four times more bioavailable via i.p. injection than via the oral route of administration. This may account for the increased hypoglycemic effect seen in the i.p. glucose tolerance test vs that seen orally. Our results suggest that puerarin is rapidly absorbed from the intestine without metabolism, while daidzin is hydrolyzed to the aglycone daidzein. The opposing effects of puerarin and daidzin on glucose homeostasis may have implications for the activity of dietary supplements that contain both of these isoflavonoids.     

Quercetin exhibits a specific fluorescence in cellular milieu: a valuable tool for the study of its intracellular distribution

The elaboration of novel techniques for flavonoid intracellular tracing would elucidate the compounds' absorption and bioavailability and assist molecular and pharmacological approaches, as they are promising candidates for drug development. This study exploited the properties of quercetin (3,3',4',5,7-pentahydroxyflavone), found in high concentrations in the majority of edible plants. Through the use of UV-vis spectroscopy, confocal microscopy, and HPLC-ESI-MS, native quercetin, at physiologically relevant concentrations, was found to exhibit a specific fluorescence (488 nmex/500-540 nmem) upon internalization. This fluorescence shift is due to a non-covalent binding to intracellular targets (probably proteins) and compatible with the settings applied in confocal microscopy. This property provides a valuable, selective alternative technique for quercetin tracing in cellular systems, permitting the quantitative evaluation of its transit at pharmacologically relevant concentrations and the validation of a number of already described molecular functions.     

Soil CO2 concentration,efflux, and partitioning in a recently afforested grassland

Relatively few studies have documented the impacts of afforestation, particularly production forestry, on belowground carbon dioxide (CO₂) effluxes to the atmosphere. We evaluated the changes in the soil CO₂ efflux—a proxy for soil respiration (Rs)—for three years following a native grassland conversion to eucalypt plantations in southern Brazil where minimum tillage during site preparation created two distinct soil zones, within planting row (W) and between-row (B). We used root-exclusion and carbon (C)- isotopic approaches to distinguish Rs components (heterotrophic-Rh and autotrophic-Ra respirations), and a CO₂ profile tube (1-m deep) to determine the concentration ([CO₂]) and isotopic C signature of soil CO₂ (δ¹³[CO₂]). The soil CO₂ efflux in the afforested site averaged 0.37 g CO₂ m^?2 h^?1, which was 56% lower than the soil CO₂ efflux in the grassland. The δ¹³CO₂ in the afforested site ranged from ? 14.1‰ to ? 29.4‰, indicating a greater contribution of eucalypt-derived respiration (both Rh and Ra) over time. Higher soil CO₂ efflux and lower [CO₂] were observed in W than B, indicating that soil preparation creates two distinct soil functional zones with respect to C cycling. The [CO₂] and δ¹³[CO₂] decreased in both zonal positions with eucalypt stand development. Although the equilibrium in C fluxes and pools across multiple rotations is needed to fully account for the feedback of eucalypt planted forests to climate change, we provide quantitative information on soil CO₂ dynamics after afforestation and show how soil preparation can leverage the feedback of planted forests to climate change.

     

Effects of environment on compensatory mutations to ameliorate costs of antibiotic resistance

Most types of antibiotic resistance impose a biological cost on bacterial fitness. These costs can be compensated, usually without loss of resistance, by second-site mutations during the evolution of the resistant bacteria in an experimental host or in a laboratory medium. Different fitness-compensating mutations were selected depending on whether the bacteria evolved through serial passage in mice or in a laboratory medium. This difference in mutation spectra was caused by either a growth condition-specific formation or selection of the compensated mutants. These results suggest that bacterial evolution to reduce the costs of antibiotic resistance can take different trajectories within and outside a host.     

A heat-sensitive TRP channel expressed in keratinocytes

Mechanical and thermal cues stimulate a specialized group of sensory neurons that terminate in the skin. Three members of the transient receptor potential (TRP) family of channels are expressed in subsets of these neurons and are activated at distinct physiological temperatures. Here, we describe the cloning and characterization of a novel thermosensitive TRP channel. TRPV3 has a unique threshold: It is activated at innocuous (warm) temperatures and shows an increased response at noxious temperatures. TRPV3 is specifically expressed in keratinocytes; hence, skin cells are capable of detecting heat via molecules similar to those in heat-sensing neurons.     

Chlorpromazine and its metabolites alter polymerization and gelation of actin

Hepatic hydroxylated metabolites of chlorpromazine (10(-5)M to 10(-4)M), a frequently used phenothiazine tranquilizer, produce solid gel formation with filamentous actin, but the less toxic chlorpromazine sulfoxide metabolite does not. At higher concentrations (5 x 10(-4)M) chlorpromazine inhibits actin polymerization. These dose-response relationships parallel the drug's hepatic toxicity in vivo and suggest that interactions between chloropromazine or chlorpromazine metabolites and actin could be an underlying mechanism of cell injury.     

Time of emergence of Rumex crispus L. as affected by dispersal time,soil cover,and mechanical disturbance

Abstract

Emergence pattern of the perennial weed species Rumex crispus L. was studied under semi-field conditions. Seeds from three populations were harvested from the mother plants and sown in pots buried in an experimental field, either in late autumn or after winter storage at outdoor temperatures. Seeds were sown on the soil surface or covered by a 2-cm soil layer. In addition, some of the seeds sown in autumn were also subjected to mechanical disturbance in autumn or in spring. Population and soil cover both had a strong effect on total emergence. The light requirement of the species for germination was indicated by a higher rate of emergence from seeds sown on the soil surface than from covered seeds. Sowing date influenced the timing of emergence but not the total emergence. Autumn sowing led to earlier and more concentrated emergence, while seedlings from spring-sown seeds showed a more intermittent emergence pattern. Stirring after sowing had a positive effect on emergence compared with emergence from undisturbed, covered seeds. In all populations and treatments, emergence continued throughout the growing season, contradicting earlier findings that R. crispus seeds germinate mainly in early spring and autumn, and enter secondary dormancy during summer. The delay of emergence in seeds that had remained on the mother plant over winter indicates a certain level of dormancy, which was gradually broken in early summer.